Anti-pd-1 antibody for use in a method of treatment of recurrent small cell lung cancer

ABSTRACT

This disclosure provides a method for treating a subject afflicted with a tumor derived from a small cell lung cancer, which method comprises administering to the subject an antibody or an antigen-binding portion thereof that specifically binds to a Programmed Death-1 (PD-1) receptor and inhibits PD-1 activity or the combination of (a) an antibody or an antigen-binding portion thereof that specifically binds to a PD-1 receptor and inhibits PD-1 activity; and (b) an antibody or an antigen-binding portion thereof that specifically binds to a Cytotoxic T-Lymphocyte Antigen-4 (CTLA-4) and inhibits CTLA-4 activity.

FIELD OF THE DISCLOSURE

This disclosure relates to methods for treating a tumor derived from asmall cell lung cancer in a subject comprising administering to thesubject an anti-Programmed Death-1 (PD-1) antibody or a combination ofan anti-PD-1 antibody and an anti-Cytotoxic T-Lymphocyte Antigen-4(CTLA-4) antibody.

BACKGROUND OF THE DISCLOSURE

Human cancers harbor numerous genetic and epigenetic alterations,generating neoantigens potentially recognizable by the immune system(Sjoblom et al., (2006) Science 314:268-74). The adaptive immune system,comprised of T and B lymphocytes, has powerful anti-cancer potential,with a broad capacity and exquisite specificity to respond to diversetumor antigens. Further, the immune system demonstrates considerableplasticity and a memory component. The successful harnessing of allthese attributes of the adaptive immune system would make immunotherapyunique among all cancer treatment modalities.

PD-1 is a key immune checkpoint receptor expressed by activated T and Bcells and mediates immunosuppression. PD-1 is a member of the CD28family of receptors, which includes CD28, CTLA-4, ICOS, PD-1, and BTLA.Two cell surface glycoprotein ligands for PD-1 have been identified,Programmed Death Ligand-1 (PD-L1) and Programmed Death Ligand-2 (PD-L2),that are expressed on antigen-presenting cells as well as many humancancers and have been shown to down-regulate T cell activation andcytokine secretion upon binding to PD-1.

Nivolumab (formerly designated 5C4, BMS-936558, MDX-1106, or ONO-4538)is a fully human IgG4 (S228P) PD-1 immune checkpoint inhibitor antibodythat selectively prevents interaction with PD-1 ligands (PD-L1 andPD-L2), thereby blocking the down-regulation of antitumor T-cellfunctions (U.S. Pat. No. 8,008,449; Wang et al., 2014 Cancer ImmunolRes. 2(9):846-56).

Ipilimumab (YERVOY®) is a fully human, IgG1 monoclonal antibody thatblocks the binding of CTLA-4 to its B7 ligands, thereby stimulating Tcell activation and improving overall survival (OS) in patients withadvanced melanoma (Hodi et al. (2010) N Engl J Med 363:711-23).Concurrent therapy with nivolumab and ipilimumab in a Phase 1 clinicaltrial produced rapid and deep tumor regression in a substantialproportion of patients with advanced melanoma, and was significantlymore effective than either antibody alone (Wolchok et al. (2013) N EnglJ Med 369(2):122-33; WO 2013/173223). However, it was hitherto not knownwhether this combination of immunoregulatory antibodies would besimilarly effective for the treatment of small-cell lung cancer (SCLC).

SCLC accounts for approximately 14% of all lung cancers (Byers andRudin, Cancer 121:664-72 (2015)). Most patients present withextensive-stage disease characterized by widespread metastases and poorsurvival. Although 35% to 86% of patients respond to first-linechemotherapy, disease progresses rapidly, and outcomes with second-linetreatment are poor (Hanna et al., J. Clin. Oncol. 24:2038-43 (2006);Puglisi et al., Br. J. Cancer 102:629-38 (2010); Zatloukal et al., Ann.Oncol. 21:1810-16 (2010)). Accordingly, there remains a need foreffective therapies for the treatment of SCLC and, in particular, secondline therapies for patients that relapse following initial treatment.

SUMMARY OF THE DISCLOSURE

The present disclosure relates to a method for treating a subjectafflicted with a tumor derived from a small-cell lung cancer (SCLC)comprising administering to the subject a combination of: (a) anantibody or an antigen-binding portion thereof that binds specificallyto a Programmed Death-1 (PD-1) receptor and inhibits PD-1 activity(“anti-PD-1 antibody”).

Other aspects of the present disclosure relate to a method for treatinga subject afflicted with a tumor derived from a SCLC comprisingadministering to the subject a combination of: (a) an anti-PD-1 antibodyand (b) an antibody or an antigen-binding portion thereof that bindsspecifically to Cytotoxic T-Lymphocyte Antigen-4 (CTLA-4) and inhibitsCTLA-4 activity (“anti-CTLA-4 antibody”).

In some embodiments, the anti-PD-1 antibody cross-competes withnivolumab for binding to human PD-1. In embodiments, the anti-PD-1antibody is a chimeric, humanized or human monoclonal antibody or aportion thereof. In other embodiments, the anti-PD-1 antibody comprisesa heavy chain constant region which is of a human IgG1 or IgG4 isotype.In certain embodiment, the anti-PD-1 antibody is nivolumab. In oneembodiment, the anti-PD-1 antibody is pembrolizumab.

In some embodiments, the anti-CTLA-4 antibody is a chimeric, humanizedor human monoclonal antibody or a portion thereof. In some embodiments,the anti-CTLA-4 antibody comprises a heavy chain constant region whichis of a human IgG1 isotype. In certain embodiments, the anti-CTLA-4antibody is ipilimumab. In other embodiments, the anti-CTLA-4 antibodyis tremelimumab. In some embodiments, the anti-CTLA-4 antibody ocross-competes with ipilimumab for binding to human CTLA-4.

In some embodiments, the anti-PD-1 antibody or antigen-binding portionthereof is administered at a dose of about 1 mg/kg or about 3 mg/kg bodyweight once about every 2 weeks. In other embodiments, the anti-PD-1antibody or antigen-binding portion thereof is administered at a dose ofabout 1 mg/kg or about 3 mg/kg body weight once about every 3 weeks. Incertain embodiments, the anti-CTLA-4 antibody or antigen-binding portionthereof is administered at a dose of about 1 mg/kg or about 3 mg/kg bodyweight. In other embodiments, the anti-PD-1 antibody or antigen-bindingportion thereof is administered at a dose of about 3 mg/kg body weightonce about every 3 weeks and the anti-CTLA-4 antibody or antigen-bindingportion thereof is administered at a dose of about 1 mg/kg body weightonce about every 3 weeks. In certain embodiments, the anti-PD-1 antibodyor antigen-binding portion thereof is administered at a dose of about 1mg/kg body weight once about every 3 weeks and the anti-CTLA-4 antibodyor antigen-binding portion thereof is administered at a dose of about 3mg/kg body weight once about every 3 weeks.

In certain embodiments, a subject treated with a disclosed methodexhibits progression-free survival of at least about one month, at leastabout 2 months, at least about 3 months, at least about 4 months, atleast about 5 months, at least about 6 months, at least about 7 months,at least about 8 months, at least about 9 months, at least about 10months, at least about 11 months, at least about one year, at leastabout eighteen months, at least about two years, at least about threeyears, at least about four years, or at least about five years. In someembodiments, the subject exhibits progression-free survival of at leastabout eight months after the initial administration.

In embodiments, the subject has a lung tumor that has ≥1% PD-L1expression. In certain embodiments, the anti-PD-1 antibody or theanti-PD-1 antibody and anti-CTLA-4 combination is administered for aslong as clinical benefit is observed or until disease progression orunmanageable toxicity occurs. In one embodiment, the anti-PD-1 and/oranti-CTLA-4 antibodies are formulated for intravenous administration. Incertain embodiments, the anti PD-1 antibody and the anti-CTLA-4 antibodyare administered sequentially to the subject. In some embodiments, theanti-PD-1 and anti-CTLA-4 antibodies are administered within 30 minutesof each other. In one embodiment, the anti-PD-1 antibody is administeredbefore the anti-CTLA-4 antibody. In another embodiment, the anti-CTLA-4antibody is administered before the anti-PD-1 antibody. In someembodiments, the anti-PD-1 antibody and the anti-CTLA-4 antibody areadministered concurrently in separate compositions. In certainembodiments, the anti-PD-1 antibody and the anti-CTLA-4 antibody areadministered concurrently as a single composition.

In one embodiment, the anti-PD-1 antibody is administered at asubtherapeutic dose. In certain embodiments, the anti-CTLA-4 antibody.In some embodiments, the anti-PD-1 antibody and the anti-CTLA-4 antibodyare each administered at a subtherapeutic dose.

The present disclosure further relates to a kit for treating a subjectafflicted with a tumor derived from a SCLC, the kit comprising: (a) anamount ranging from about 4 mg to about 500 mg of an anti-PD-1 antibody;and (b) instructions for using the PD-1 antibody and the CTLA-4 antibodyin any disclosed method.

The present disclosure further relates to a kit for treating a subjectafflicted with a tumor derived from a SCLC, the kit comprising: (a) anamount ranging from about 4 mg to about 500 mg of an anti-PD-1 antibody;(b) an amount ranging from about 4 mg to about 500 mg of a CTLA-4antibody; and (c) instructions for using the PD-1 antibody and theCTLA-4 antibody in any disclosed method.

Other features and advantages of the instant disclosure will be apparentfrom the following detailed description and examples which should not beconstrued as limiting. The contents of all cited references, includingscientific articles, newspaper reports, GenBank entries, patents andpatent applications cited throughout this application are expresslyincorporated herein by reference.

Embodiments

E1. A method for treating a subject afflicted with a tumor derived froma small-cell lung cancer (SCLC) comprising administering to the subjectan antibody or an antigen-binding portion thereof that bindsspecifically to a Programmed Death-1 (PD-1) receptor and inhibits PD-1activity (“anti-PD-1 antibody”).

E2. The method of embodiment E1, further comprising administering to thesubject an antibody or an antigen-binding portion thereof that bindsspecifically to Cytotoxic T-Lymphocyte Antigen-4 (CTLA-4) and inhibitsCTLA-4 activity (“anti-CTLA-4 antibody”).

E3. The method of embodiment E1 or E2, wherein the SCLC comprises asmall cell carcinoma.

E4. The method of embodiment E1 or E2, wherein the SCLC comprises acombined small cell carcinoma.

E5. The method of any one of embodiments E1 to E4, wherein the SCLC is arecurrent SCLC.

E6. The method of any one of embodiments E1 to E5, wherein the subjectreceived at least one, at least two, at least three, at least four, orat least five previous lines of therapy to treat the tumor.

E7. The method of embodiment E6, wherein the previous line of therapycomprises a chemotherapy.

E8. The method of embodiment E7, wherein the chemotherapy comprises aplatinum-based therapy.

E9. The method of embodiment E8, wherein the platinum-based therapycomprises a platinum-based antineoplastic selected from the groupconsisting of cisplatin, carboplatin, oxaliplatin, nedaplatin, triplatintetranitrate, phenanthriplatin, picoplatin, satraplatin, and anycombination thereof.

E10. The method of embodiment E7 or E8, wherein the platinum-basedtherapy comprises cisplatin.

E11. The method of any one of embodiments E1 to E10, wherein theanti-PD-1 antibody cross-competes with nivolumab for binding to humanPD-1.

E12. The method of any one of embodiments E1 to E11, wherein theanti-PD-1 antibody binds to the same epitope as nivolumab.

E13. The method of any one of embodiments E1 to E12, wherein theanti-PD-1 antibody is a chimeric, humanized or human monoclonal antibodyor a portion thereof.

E14. The method of any one of embodiments E1 to E13, wherein theanti-PD-1 antibody comprises a heavy chain constant region which is of ahuman IgG1 or IgG4 isotype.

E15. The method of any one of embodiments E1 to E14, wherein theanti-PD-1 antibody is nivolumab.

E16. The method of any one of embodiments E1 to E14, wherein theanti-PD-1 antibody is pembrolizumab.

E17. The method of any one of embodiments E2 to E16, wherein theanti-CTLA-4 antibody is a chimeric, humanized or human monoclonalantibody or a portion thereof.

E18. The method of any one of embodiments E2 to E17, wherein theanti-CTLA-4 antibody comprises a heavy chain constant region which is ofa human IgG1 isotype.

E19. The method of any one of embodiments E2 to E18, wherein theanti-CTLA-4 antibody is ipilimumab.

E20. The method of any one of embodiments E2 to E18, wherein theanti-CTLA-4 antibody is tremelimumab.

E21. The method of any one of embodiments E2 to E20, wherein theanti-CTLA-4 antibody cross-competes with ipilimumab for binding to humanCTLA-4.

E22. The method of any one of embodiments E1 to E21, wherein theanti-PD-1 antibody is administered at a dose ranging from at least about0.1 mg/kg to at least about 10.0 mg/kg body weight once about every 1,2, 3, or 4 weeks.

E23. The method of any one of embodiments E1 to E22, wherein theanti-PD-1 antibody is administered at a dose of about 1 mg/kg or about 3mg/kg body weight

E24. The method of any one of embodiments E1 to E21, wherein theanti-PD-1 antibody or antigen-binding portion thereof is administered ata flat dose.

E25. The method of any one of embodiments E1 to E21 and E24, wherein theanti-PD-1 antibody or antigen-binding portion thereof is administered ata flat dose of at least about 200, at least about 220, at least about240, at least about 260, at least about 280, at least about 300, atleast about 320, at least about 340, at least about 360, at least about380, at least about 400, at least about 420, at least about 440, atleast about 460, at least about 480, at least about 500 or at leastabout 550 mg.

E26. The method of any one of embodiments E1 to E21, E24, and E25,wherein the anti-PD-1 antibody or antigen-binding portion thereof isadministered at a flat dose about once every 1, 2, 3, or 4 weeks.

E27. The method of anyone of embodiments E1 to E26, wherein theanti-PD-1 antibody is administered once about every 2 weeks.

E28. The method of anyone of embodiments E1 to E26, wherein theanti-PD-1 antibody is administered once about every 3 weeks.

E29. The method of any one of embodiments E1 to E28, wherein theanti-PD-1 antibody is administered for as long as clinical benefit isobserved or until unmanageable toxicity or disease progression occurs.

E30. The method of any one of embodiments E2 to E29, wherein theanti-CTLA-4 antibody is administered at a dose ranging from at leastabout 0.1 mg/kg to at least about 10.0 mg/kg body weight once aboutevery 1, 2, 3, or 4 weeks.

E31. The method of any one of embodiments E2 to E30, wherein theanti-CTLA-4 is administered at a dose of about 1 mg/kg or about 3 mg/kgbody weight.

E32. The method of any one of embodiments E2 to E31, wherein theanti-PD-1 antibody or antigen-binding portion thereof is administered ata flat dose.

E33. The method of anyone of embodiments E2 to E32, wherein theanti-CTLA-4 antibody is administered once about every 2 weeks.

E34. The method of anyone of embodiments E2 to E32, wherein theanti-CTLA-4 antibody is administered once about every 3 weeks.

E35. The method of any one of embodiments E2 to E34, wherein theanti-PD-1 antibody is administered at a dose of about 3 mg/kg bodyweight once about every 3 weeks and the anti-CTLA-4 antibody orantigen-binding portion thereof is administered at a dose of about 1mg/kg body weight once about every 3 weeks.

E36. The method of any one of embodiments E2 to E34, wherein theanti-PD-1 antibody is administered at a dose of about 1 mg/kg bodyweight once about every 3 weeks and the anti-CTLA-4 antibody orantigen-binding portion thereof is administered at a dose of about 3mg/kg body weight once about every 3 weeks.

E37. The method of any one of embodiments E1 to E36, wherein the subjectexhibits progression-free survival of at least about one month, at leastabout 2 months, at least about 3 months, at least about 4 months, atleast about 5 months, at least about 6 months, at least about 7 months,at least about 8 months, at least about 9 months, at least about 10months, at least about 11 months, at least about one year, at leastabout eighteen months, at least about two years, at least about threeyears, at least about four years, or at least about five years after theinitial administration.

E38. The method of any one of embodiments E1 to E37, wherein the subjecthas a tumor that has ≥1% PD-L1 expression.

E39. The method of any one of embodiments E1 to E38, wherein the subjecthas a tumor that has ≥5% PD-L1 expression.

E40. The method of any one of embodiments E2 to E39, wherein thecombination is administered for as long as clinical benefit is observedor until disease progression or unmanageable toxicity occurs.

E41. The method of any one of embodiments E1 to E40, wherein theanti-PD-1 antibody is formulated for intravenous administration.

E42. The method of any one of embodiments E2 to E41, wherein theanti-CTLA-4 antibody is formulated for intravenous administration.

E43. The method of any one of embodiments E2 to E42, wherein theanti-PD-1 antibody and the anti-CTLA-4 antibody are administeredsequentially to the subject.

E44. The method of any one of claims 2-43, wherein the anti-PD-1 andanti-CTLA-4 antibodies are administered within 30 minutes of each other.

E45. The method of any one of embodiments E2 to E44, wherein theanti-PD-1 antibody or antigen-binding portion thereof is administeredbefore the anti-CTLA-4 antibody or antigen-binding portion thereof.

E46. The method of any one of embodiments E2 to E44, wherein theanti-CTLA-4 antibody or antigen-binding portion thereof is administeredbefore the anti-PD-1 antibody or antigen-binding portion thereof.

E47. The method of any one of embodiments E2 to E42, wherein theanti-PD-1 antibody or antigen-binding portion thereof and theanti-CTLA-4 antibody or antigen-binding portion thereof are administeredconcurrently in separate compositions.

E48. The method of any one of embodiments E2 to E42, wherein theanti-PD-1 antibody or antigen-binding portion thereof and theanti-CTLA-4 antibody or antigen-binding portion thereof are administeredconcurrently as a single composition.

E49. The method of any one of embodiments E1 to E48, wherein theanti-PD-1 antibody or antigen-binding portion thereof is administered ata subtherapeutic dose.

E50. The method of any one of embodiments E2 to E49, wherein theanti-CTLA-4 antibody or antigen-binding portion thereof is administeredat a subtherapeutic dose.

E51. The method of any one of embodiments E2 to E50, wherein theanti-PD-1 antibody or antigen-binding portion thereof and theanti-CTLA-4 antibody or antigen-binding portion thereof are eachadministered at a subtherapeutic dose.

E52. A kit for treating a subject afflicted with a tumor derived from anSCLC, the kit comprising:

-   (a) an amount ranging from about 4 mg to about 500 mg of an    anti-PD-1 antibody or an antigen-binding portion thereof; and-   (b) instructions for using the PD-1 antibody in the method of any of    embodiments E1 to E51.

E53. A kit for treating a subject afflicted with a tumor derived from anSCLC, the kit comprising:

-   (a) an amount ranging from about 4 mg to about 500 mg of an    anti-PD-1 antibody or an antigen-binding portion thereof;-   (b) an amount ranging from about 4 mg to about 500 mg of a CTLA-4    antibody or an antigen-binding portion thereof; and-   (c) instructions for using the PD-1 antibody and the CTLA-4 antibody    in the method of any of embodiments E2 to E51.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows a schematic representation of a study design for treatmentof a tumor derived from an SCLC using an anti-PD-1 antibody or acombination of an anti-PD-1 antibody and an anti-CTLA-4 antibody.

FIGS. 2A and 2B show images of a tumor derived from a SCLC found in asubject's lungs before (FIG. 2A) and after (FIG. 2B) treatment with acombination of nivolumab 1 mg/kg body weight and ipilimumab 3 mg/kg bodyweight. Tumor tissue is circled in both FIG. 2A and FIG. 2B.

FIGS. 3A-3C provide graphical representations of the changes in tumorburden in individual patients receiving nivolumab 3 mg/kg (FIG. 3A),nivolumab 1 mg/kg plus ipilimumab 3 mg/kg (FIG. 3B), and nivolumab 3mg/kg plus ipilimumab 1 mg/kg (FIG. 3C). Only patients with targetlesions at baseline and with at least one on-treatment tumor assessmentwere included (nivolumab 3 mg/kg, n=80; nivolumab 1 mg/kg plusipilitnumab 3 mg/kg, n=46; nivolumab 3 mg/kg plus ipilimumab 1 mg/kg,n=47). Horizontal grey lines indicate the 30% reduction consistent witha Response Evaluation Criteria in Solid Tumors (RECIST; version 1.1)objective response. Subjects displaying a complete response (CR) or apartial response (PR) are marked by an inverted triangle at the point ofPR or CR. A plus sign indicates the first occurrence of a new lesion,and an open circle indicates the point at which the subject went offtreatment. Measurements in excess of 100% were truncated to 100%, andthe data point is marked by an open square.

FIGS. 4A and 4B provide Kaplan-Meier curves of overall survival (FIG.4A) and progression-free survival (FIG. 4B) for subjects treated withnivolumab 3 mg/kg (circles), nivolumab 1 mg/kg plus ipilimumab 3 mg/kg(diamonds), and nivolumab 3 mg/kg plus ipilimumab 1 mg/kg (triangles).The numbers of subjects at risk and the numbers of censored patients areshown for each treatment below the y-axes (FIGS. 4A and 4B).

FIGS. 5A-5C provide graphical representations of the changes in tumorburden according to tumor PD-L1 expression status. The percent changefrom baseline for tumors with less than 1% PD-L1 expression are shown ingrey and for tumors with ≥1% PD-L1 expression shown in black. The changein baseline for tumors where PD-L1 expression was not evaluable ormissing are shown in light grey. Closed circles above a bar indicate aconfirmed responder. Open boxes indicate that the % change was truncatedto 100%.

FIGS. 6A-6D provide Kaplan-Meier curves of overall survival (FIGS. 6Aand 6B) and progression-free survival (FIGS. 6C and 6D) for subjectswith one prior therapy (FIG. 6A and FIG. 6C) and two or more priortherapies (FIG. 6B and FIG. 6D). Subjects were treated with nivolumab 3mg/kg (circles), nivolumab 1 mg/kg plus ipilimumab 3 mg/kg (diamonds),or nivolumab 3 mg/kg plus ipilimumab 1 mg/kg (triangles). The numbers ofsubjects at risk and the numbers of censored patients are shown for eachtreatment below the y-axes (FIGS. 6A-6D).

FIGS. 7A-7C provide graphical representations of the changes in tumorburden according to platinum sensitivity in individual patientsreceiving nivolumab 3 mg/kg (FIG. 7A), nivolumab 1 mg/kg plus ipilimumab3 mg/kg (FIG. 7B), and nivolumab 3 mg/kg plus ipilimumab 1 mg/kg (FIG.7C) (platinum sensitive tumors are indicated by grey lines and platinumresistant tumors are indicated by black lines). Only patients withtarget lesions at baseline and with ≥1 on-treatment tumor assessmentwere included (nivolumab-3, n=31; nivolumab-1/ipilimumab-3, n=21;nivolumab-3/ipilimumab-1, n=17). Panels show the tumor burden (assessedas the longest linear dimension) over time in patients receivingsecond-line nivolumab-3 (FIG. 7A), nivolumab-1/ipilimumab-3 (FIG. 7B),and nivolumab-3/ipilimumab-1 (FIG. 7C). Horizontal reference lineindicates the 30% reduction consistent with a RECIST (version 1.1)objective response. Subjects displaying a complete response (CR) or apartial response (PR) are marked by an inverted triangle at the point ofPR or CR. A plus sign indicates the first occurrence of a new lesion,and an open circle indicates the point at which the subject went offtreatment. Measurements in excess of 100% were truncated to 100%, andthe data point is marked by an open square.

FIG. 8 shows a schematic representation of a study design for treatmentof a tumor derived from an SCLC using an anti-PD-1 antibody or acombination of an anti-PD-1 antibody and an anti-CTLA-4 antibody, whichincludes a randomized cohort and a non-randomized cohort.

FIG. 9 provides a Kaplan-Meier curve of overall survival for subjectstreated with nivolumab 3 mg/kg (circles) or nivolumab 1 mg/kg plusipilimumab 3 mg/kg (diamonds). The numbers of subjects at risk are shownfor each treatment below the y-axis.

FIG. 10 provides a graphical representation of the overall response rate(ORR) observed in patients treated with nivolumab 1 mg/kg or nivolumab 1mg/kg plus ipilimumab 3 mg/kg, in the randomized cohort and thenon-randomized cohort. Nivo=nivolumab; ipi=ipilimumab.

FIG. 11 provides a graphical representation of the rate of progressionfree survival (PFS) observed in patients treated with nivolumab 3 mg/kgor nivolumab 1 mg/kg plus ipilimumab 3 mg/kg, in the randomized cohortand the non-randomized cohort. Nivo=nivolumab; ipi=ipilimumab.

FIG. 12 provides a graphical representation of the overall survival (OS)rate observed in patients treated with nivolumab 3 mg/kg or nivolumab 1mg/kg plus ipilimumab 3 mg/kg, in the randomized cohort and thenon-randomized cohort. Nivo=nivolumab; ipi=ipilimumab.

DETAILED DESCRIPTION OF THE DISCLOSURE

The present disclosure relates to methods for treating a tumor derivedfrom an SCLC in a patient comprising administering to the patient ananti-PD-1 antibody or a combination of an anti-PD-1 antibody and ananti-CTLA-4 antibody.

Terms

In order that the present disclosure can be more readily understood,certain terms are first defined. As used in this application, except asotherwise expressly provided herein, each of the following terms shallhave the meaning set forth below. Additional definitions are set forththroughout the application.

“Administering” refers to the physical introduction of a compositioncomprising a therapeutic agent to a subject, using any of the variousmethods and delivery systems known to those skilled in the art. Routesof administration for the anti-PD-1 antibody include intravenous,intramuscular, subcutaneous, intraperitoneal, spinal, or otherparenteral routes of administration, for example by injection orinfusion. The phrase “parenteral administration” as used herein meansmodes of administration other than enteral and topical administration,usually by injection, and includes, without limitation, intravenous,intramuscular, intraarterial, intrathecal, intralymphatic,intralesional, intracapsular, intraorbital, intracardiac, intradermal,intraperitoneal, transtracheal, subcutaneous, subcuticular,intraarticular, subcapsular, subarachnoid, intraspinal, epidural andintrasternal injection and infusion, as well as in vivo electroporation.In some embodiments, the combination is administered via anon-parenteral route, in some embodiments, orally. Other non-parenteralroutes include a topical, epidermal or mucosal route of administration,for example, intranasally, vaginally, rectally, sublingually ortopically. Administering can also be performed, for example, once, aplurality of times, and/or over one or more extended periods.

An “adverse event” (AE) as used herein is any unfavorable and generallyunintended or undesirable sign (including an abnormal laboratoryfinding), symptom, or disease associated with the use of a medicaltreatment. For example, an adverse event can be associated withactivation of the immune system or expansion of immune system cells(e.g., T cells) in response to a treatment. A medical treatment can haveone or more associated AEs and each AE can have the same or differentlevel of severity. Reference to methods capable of “altering adverseevents” means a treatment regime that decreases the incidence and/orseverity of one or more AEs associated with the use of a differenttreatment regime.

An “antibody” (Ab) shall include, without limitation, a glycoproteinimmunoglobulin which binds specifically to an antigen and comprises atleast two heavy (H) chains and two light (L) chains interconnected bydisulfide bonds, or an antigen-binding portion thereof. Each H chaincomprises a heavy chain variable region (abbreviated herein as V_(H))and a heavy chain constant region. The heavy chain constant regioncomprises at least three constant domains, C_(H1), C_(H2) and C_(H3).Each light chain comprises a light chain variable region (abbreviatedherein as V_(L)) and a light chain constant region. The light chainconstant region is comprises one constant domain, C_(L). The V_(H) andV_(L) regions can be further subdivided into regions ofhypervariability, termed complementarity determining regions (CDRs),interspersed with regions that are more conserved, termed frameworkregions (FRs). Each V_(H) and V_(L) comprises three CDRs and four FRs,arranged from amino-terminus to carboxy-terminus in the following order:FR1, CDR1, FR2, CDR2, FR3, CDR3, and FR4. The variable regions of theheavy and light chains contain a binding domain that interacts with anantigen. The constant regions of the antibodies can mediate the bindingof the immunoglobulin to host tissues or factors, including variouscells of the immune system (e.g., effector cells) and the firstcomponent (C1q) of the classical complement system.

An immunoglobulin can derive from any of the commonly known isotypes,including but not limited to IgA, secretory IgA, IgG, and IgM. IgGsubclasses are also well known to those in the art and include but arenot limited to human IgG1, IgG2, IgG3, and IgG4. “Isotype” refers to theantibody class or subclass (e.g., IgM or IgG1) that is encoded by theheavy chain constant region genes. The term “antibody” includes, by wayof example, both naturally occurring and non-naturally occurringantibodies; monoclonal and polyclonal antibodies; chimeric and humanizedantibodies; human or non-human antibodies; wholly synthetic antibodies;and single chain antibodies. A non-human antibody can be humanized byrecombinant methods to reduce its immunogenicity in man. Where notexpressly stated, and unless the context indicates otherwise, the term“antibody” also includes an antigen-binding fragment or anantigen-binding portion of any of the aforementioned immunoglobulins,and includes a monovalent and a divalent fragment or portion, and asingle chain antibody.

An “isolated antibody” refers to an antibody that is substantially freeof other antibodies having different antigenic specificities (e.g., anisolated antibody that binds specifically to PD-1 is substantially freeof antibodies that bind specifically to antigens other than PD-1). Anisolated antibody that binds specifically to PD-1 can, however, havecross-reactivity to other antigens, such as PD-1 molecules fromdifferent species. Moreover, an isolated antibody can be substantiallyfree of other cellular material and/or chemicals.

The term “monoclonal antibody” (mAb) refers to a non-naturally occurringpreparation of antibody molecules of single molecular composition, i.e.,antibody molecules whose primary sequences are essentially identical,and which exhibits a single binding specificity and affinity for aparticular epitope. A monoclonal antibody is an example of an isolatedantibody. MAbs can be produced by hybridoma, recombinant, transgenic, orother techniques known to those skilled in the art.

A “human antibody” (HuMAb) refers to an antibody having variable regionsin which both the FRs and CDRs are derived from human germlineimmunoglobulin sequences. Furthermore, if the antibody contains aconstant region, the constant region also is derived from human germlineimmunoglobulin sequences. The human antibodies of the disclosure caninclude amino acid residues not encoded by human germline immunoglobulinsequences (e.g., mutations introduced by random or site-specificmutagenesis in vitro or by somatic mutation in vivo). However, the term“human antibody,” as used herein, is not intended to include antibodiesin which CDR sequences derived from the germline of another mammalianspecies, such as a mouse, have been grafted onto human frameworksequences. The terms “human antibodies” and “fully human antibodies” areused synonymously.

A “humanized antibody” refers to an antibody in which some, most or allof the amino acids outside the CDRs of a non-human antibody are replacedwith corresponding amino acids derived from human immunoglobulins. Inone embodiment of a humanized form of an antibody, some, most or all ofthe amino acids outside the CDRs have been replaced with amino acidsfrom human immunoglobulins, whereas some, most, or all amino acidswithin one or more CDRs are unchanged. Small additions, deletions,insertions, substitutions, or modifications of amino acids arepermissible as long as they do not abrogate the ability of the antibodyto bind to a particular antigen. A “humanized antibody” retains anantigenic specificity similar to that of the original antibody. In someembodiments, the CDRs of a humanized antibody contain CDRs from anon-human, mammalian antibody. In other embodiments, the CDRs of ahumanized antibody contain CDRs from an engineered, synthetic antibody.

A “chimeric antibody” refers to an antibody in which the variableregions are derived from one species and the constant regions arederived from another species, such as an antibody in which the variableregions are derived from a mouse antibody and the constant regions arederived from a human antibody.

An “anti-antigen antibody” refers to an antibody that binds specificallyto the antigen. For example, an anti-PD-1 antibody binds specifically toPD-1 and an anti-CTLA-4 antibody binds specifically to CTLA-4.

An “antigen-binding portion” of an antibody (also called an“antigen-binding fragment”) refers to one or more fragments of anantibody that retain the ability to bind specifically to the antigenbound by the whole antibody.

A “cancer” refers a broad group of various diseases characterized by theuncontrolled growth of abnormal cells in the body. A “cancer” or “cancertissue” can include a tumor. Unregulated cell division and growthresults in the formation of malignant tumors that invade neighboringtissues and may also metastasize to distant parts of the body throughthe lymphatic system or bloodstream. Following metastasis, the distaltumors can be said to be “derived from” the original, pre-metastasistumor. For example, a “tumor derived from” an SCLC refers to a tumorthat is the result of a metastasized SCLC. Because the distal tumor isderived from the pre-metastasis tumor, the “derived from” tumor can alsocomprise the pre-metastasis tumor, e.g., a tumor derived from an SCLCcan comprise an SCLC.

“Cytotoxic T-Lymphocyte Antigen-4” (CTLA-4) refers to animmunoinhibitory receptor belonging to the CD28 family. CTLA-4 isexpressed exclusively on T cells in vivo, and binds to two ligands, CD80and CD86 (also called B7-1 and B7-2, respectively). The term “CTLA-4” asused herein includes human CTLA-4 (hCTLA-4), variants, isoforms, specieshomologs of hCTLA-4, and analogs having at least one common epitope withhCTLA-4. The complete hCTLA-4 sequence can be found under GenBankAccession No. AAB59385.

The term “immunotherapy” refers to the treatment of a subject afflictedwith, or at risk of contracting or suffering a recurrence of, a diseaseby a method comprising inducing, enhancing, suppressing or otherwisemodifying an immune response. “Treatment” or “therapy” of a subjectrefers to any type of intervention or process performed on, or theadministration of an active agent to, the subject with the objective ofreversing, alleviating, ameliorating, inhibiting, slowing down orpreventing the onset, progression, development, severity or recurrenceof a symptom, complication or condition, or biochemical indiciaassociated with a disease.

“PD-L1 positive” as used herein can be interchangeably used with “PD-L1expression of at least about 1%.” In one embodiment, the PD-L1expression can be used by any methods known in the art. In anotherembodiment, the PD-L1 expression is measured by an automated IHC. PD-L1positive tumor can thus have at least about 1%, at least about 2%, atleast about 5%, at least about 10%, at least about 20%, at least about25%, at least about 30%, at least about 40%, at least about 50%, atleast about 60%, at least about 70%, at least about 75%, at least about80%, at least about 85%, at least about 90%, at least about 95%, orabout 100% of tumor cells expressing PD-L1 as measured by an automatedIHC. In certain embodiments, “PD-L1 positive” means that there are atleast 100 cells that express PD-L1 on the surface of the cells. In otherembodiments, “PD-L2 positive” means that there are at least 100 cellsthat express PD-L2 on the surface of the cells.

“Programmed Death-1” (PD-1) refers to an immunoinhibitory receptorbelonging to the CD28 family. PD-1 is expressed predominantly onpreviously activated T cells in vivo, and binds to two ligands, PD-L1and PD-L2. The term “PD-1” as used herein includes human PD-1 (hPD-1),variants, isoforms, and species homologs of hPD-1, and analogs having atleast one common epitope with hPD-1. The complete hPD-1 sequence can befound under GenBank Accession No. U64863.

“Programmed Death Ligand-1” (PD-L1) is one of two cell surfaceglycoprotein ligands for PD-1 (the other being PD-L2) that downregulateT cell activation and cytokine secretion upon binding to PD-1. The term“PD-L1” as used herein includes human PD-L1 (hPD-L1), variants,isoforms, and species homologs of hPD-L1, and analogs having at leastone common epitope with hPD-L1. The complete hPD-L1 sequence can befound under GenBank Accession No. Q9NZQ7.

A “subject” includes any human or non-human animal. The term “non-humananimal” includes, but is not limited to, vertebrates such as non-humanprimates, sheep, dogs, and rodents such as mice, rats, and guinea pigs.In some embodiments, the subject is a human. The terms, “subject” and“patient” are used interchangeably herein.

A “therapeutically effective amount” or “therapeutically effectivedosage” of a drug or therapeutic agent is any amount of the drug that,when used alone or in combination with another therapeutic agent,protects a subject against the onset of a disease or promotes diseaseregression evidenced by a decrease in severity of disease symptoms, anincrease in frequency and duration of disease symptom-free periods, or aprevention of impairment or disability due to the disease affliction.The ability of a therapeutic agent to promote disease regression can beevaluated using a variety of methods known to the skilled practitioner,such as in human subjects during clinical trials, in animal modelsystems predictive of efficacy in humans, or by assaying the activity ofthe agent in in vitro assays.

As used herein, “subtherapeutic dose” means a dose of a therapeuticcompound (e.g., an antibody) that is lower than the usual or typicaldose of the therapeutic compound when administered alone for thetreatment of a hyperproliferative disease (e.g., cancer).

By way of example, an “anti-cancer agent” promotes cancer regression ina subject or prevents further tumor growth. In certain embodiments, atherapeutically effective amount of the drug promotes cancer regressionto the point of eliminating the cancer. “Promoting cancer regression”means that administering an effective amount of the drug, alone or incombination with an anti-neoplastic agent, results in a reduction intumor growth or size, necrosis of the tumor, a decrease in severity ofat least one disease symptom, an increase in frequency and duration ofdisease symptom-free periods, or a prevention of impairment ordisability due to the disease affliction. In addition, the terms“effective” and “effectiveness” with regard to a treatment includes bothpharmacological effectiveness and physiological safety. Pharmacologicaleffectiveness refers to the ability of the drug to promote cancerregression in the patient. Physiological safety refers to the level oftoxicity, or other adverse physiological effects at the cellular, organand/or organism level (adverse effects) resulting from administration ofthe drug.

By way of example for the treatment of tumors, a therapeuticallyeffective amount of an anti-cancer agent can inhibit cell growth ortumor growth by at least about 10%, by at least about 20%, by at leastabout 30%, by at least about 40%, by at least about 50%, by at leastabout 60%, by at least about 70%, by at least about 80%, by at leastabout 90%, by at least about 95%, or by at least about 100% relative tountreated subjects. In other embodiments of the disclosure, tumorregression can be observed and continue for a period of at least about20 days, at least about 30 days, at least about 40 days, at least about50 days, or at least about 60 days. Notwithstanding these ultimatemeasurements of therapeutic effectiveness, evaluation ofimmunotherapeutic drugs must also make allowance for “immune-relatedresponse patterns”.

An “immune-related response pattern” refers to a clinical responsepattern often observed in cancer patients treated with immunotherapeuticagents that produce antitumor effects by inducing cancer-specific immuneresponses or by modifying native immune processes. This response patternis characterized by a beneficial therapeutic effect that follows aninitial increase in tumor burden or the appearance of new lesions, whichin the evaluation of traditional chemotherapeutic agents would beclassified as disease progression and would be synonymous with drugfailure. Accordingly, proper evaluation of immunotherapeutic agents canrequire long-term monitoring of the effects of these agents on thetarget disease.

A therapeutically effective amount of a drug includes a“prophylactically effective amount,” which is any amount of the drugthat, when administered alone or in combination with an anti-neoplasticagent to a subject at risk of developing a cancer (e.g., a subjecthaving a pre-malignant condition) or of suffering a recurrence ofcancer, inhibits the development or recurrence of the cancer. In certainembodiments, the prophylactically effective amount prevents thedevelopment or recurrence of the cancer entirely. “Inhibiting” thedevelopment or recurrence of a cancer means either lessening thelikelihood of the cancer's development or recurrence, or preventing thedevelopment or recurrence of the cancer entirely.

The term “weight based dose”, as referred to herein, means that a doseadministered to a patient is calculated based on the weight of thepatient. For example, when a patient with 60 kg body weight requires 3mg/kg of an anti-PD-1 antibody, one can calculate and use theappropriate amount of the anti-PD-1 antibody (i.e., 180 mg) foradministration.

The use of the term “fixed dose” with regard to a method of thedisclosure means that two or more different antibodies in a singlecomposition (e.g., anti-PD-1 antibody and anti-CTLA-4 antibody) arepresent in the composition in particular (fixed) ratios with each other.In some embodiments, the fixed dose is based on the weight (e.g., mg) ofthe antibodies. In certain embodiments, the fixed dose is based on theconcentration (e.g., mg/ml) of the antibodies. In some embodiments, theratio is at least about 1:1, about 1:2, about 1:3, about 1:4, about 1:5,about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15,about 1:20, about 1:30, about 1:40, about 1:50, about 1:60, about 1:70,about 1:80, about 1:90, about 1:100, about 1:120, about 1:140, about1:160, about 1:180, about 1:200, about 200:1, about 180:1, about 160:1,about 140:1, about 120:1, about 100:1, about 90:1, about 80:1, about70:1, about 60:1, about 50:1, about 40:1, about 30:1, about 20:1, about15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1,about 4:1, about 3:1, or about 2:1 mg first antibody (e.g., anti-PD-1antibody) to mg second antibody (e.g., anti-CTLA-4 antibody). Forexample, the 3:1 ratio of an anti-PD-1 antibody and an anti-CTLA-4antibody can mean that a vial can contain about 240 mg of the anti-PD-1antibody and 80 mg of the anti-CTLA-4 antibody or about 3 mg/ml of theanti-PD-1 antibody and 1 mg/ml of the anti-CTLA-4 antibody.

The use of the term “flat dose” with regard to the methods and dosagesof the disclosure means a dose that is administered to a patient withoutregard for the weight or body surface area (BSA) of the patient. Theflat dose is therefore not provided as a mg/kg dose, but rather as anabsolute amount of the agent (e.g., the anti-CTLA-4 antibody and/oranti-PD-1 antibody). For example, a 60 kg person and a 100 kg personwould receive the same dose of an antibody (e.g., 240 mg of an anti-PD-1antibody).

The use of the alternative (e.g., “or”) should be understood to meaneither one, both, or any combination thereof of the alternatives. Asused herein, the indefinite articles “a” or “an” should be understood torefer to “one or more” of any recited or enumerated component.

The terms “about” or “comprising essentially of” refer to a value orcomposition that is within an acceptable error range for the particularvalue or composition as determined by one of ordinary skill in the art,which will depend in part on how the value or composition is measured ordetermined, i.e., the limitations of the measurement system. Forexample, “about” or “comprising essentially of” can mean within 1 ormore than 1 standard deviation per the practice in the art.Alternatively, “about” or “comprising essentially of” can mean a rangeof up to 10% or 20% (i.e., ±10% or ±20%). For example, about 3 mg caninclude any number between 2.7 mg and 3.3 mg (for 10%) or between 2.4 mgand 3.6 mg (for 20%). Furthermore, particularly with respect tobiological systems or processes, the terms can mean up to an order ofmagnitude or up to 5-fold of a value. When particular values orcompositions are provided in the application and claims, unlessotherwise stated, the meaning of “about” or “comprising essentially of”should be assumed to be within an acceptable error range for thatparticular value or composition.

The terms “once about every week,” “once about every two weeks,” or anyother similar dosing interval terms as used herein mean approximatenumbers. “Once about every week” can include every seven days±one day,i.e., every six days to every eight days. “Once about every two weeks”can include every fourteen days±three days, i.e., every eleven days toevery seventeen days. Similar approximations apply, for example, to onceabout every three weeks, once about every four weeks, once about everyfive weeks, once about every six weeks and once about every twelveweeks. In some embodiments, a dosing interval of once about every sixweeks or once about every twelve weeks means that the first dose can beadministered any day in the first week, and then the next dose can beadministered any day in the sixth or twelfth week, respectively. Inother embodiments, a dosing interval of once about every six weeks oronce about every twelve weeks means that the first dose is administeredon a particular day of the first week (e.g., Monday) and then the nextdose is administered on the same day of the sixth or twelfth weeks(i.e., Monday), respectively.

As described herein, any concentration range, percentage range, ratiorange or integer range is to be understood to include the value of anyinteger within the recited range and, when appropriate, fractionsthereof (such as one tenth and one hundredth of an integer), unlessotherwise indicated.

Various aspects of the disclosure are described in further detail in thefollowing subsections.

Methods of the Disclosure

This disclosure provides a method of treating a subject afflicted with atumor derived from an SCLC, which method comprises administering to thesubject an antibody or an antigen-binding portion thereof that bindsspecifically to a Programmed Death-1 (PD-1) receptor and inhibits PD-1activity (“anti-PD-1 antibody”). This disclosure further provides amethod of treating a subject afflicted with a tumor derived from anSCLC, which method comprises administering to the subject a combinationof (a) an antibody or an antigen-binding portion thereof thatspecifically binds to and a PD-1 receptor and inhibits PD-1 activity(“anti-PD-1 antibody”); and (b) an antibody or an antigen-bindingportion thereof that binds specifically to Cytotoxic T-LymphocyteAntigen-4 (CTLA-4) and inhibits CTLA-4 activity (“anti-CTLA-4antibody”). In some embodiments, the subject is a human patient.

In certain embodiments, the subject is a chemotherapy-naïve patient(e.g., a patient who has not previously received any chemotherapy). Inother embodiments, the subject has received another cancer therapy(e.g., a chemotherapy), but is resistant or refractory to such anothercancer therapy. In one particular embodiment, the SCLC is a recurrentSCLC. In some embodiments, the subject received at least one, at leasttwo, at least three, at least four, or at least five previous lines oftherapy to treat the tumor. In one embodiment, the subject received oneprevious line of therapy to treat the tumor. In another embodiment, thesubject received two previous lines of therapy to treat the tumor. Inanother embodiment, the subject received three previous lines of therapyto treat the tumor. In another embodiment, the subject received fourprevious lines of therapy to treat the tumor. In another embodiment, thesubject received five previous lines of therapy to treat the tumor. Inanother embodiment, the subject received more than five previous linesof therapy to treat the tumor.

In certain embodiments, the previous line of therapy comprises achemotherapy. In some embodiments, the chemotherapy comprises aplatinum-based therapy. In certain embodiments, the platinum basedtherapy comprises a platinum-based antineoplastic selected from thegroup consisting of cisplatin, carboplatin, oxaliplatin, nedaplatin,triplatin tetranitrate, phenanthriplatin, picoplatin, satraplatin, andany combination thereof. In one particular embodiment, theplatinum-based therapy comprises cisplatin (e.g., cisplatin incombination with etoposide). In some embodiments, the subject hasreceived a previous radiotherapy. In other embodiments, the previoustherapy comprises an antibody therapy.

In certain specific embodiments, the subject has cancer cells expressingmutated forms of the EGFR or KRAS gene. In certain embodiments, thesubject has cancer cells that are PD-L1 positive. In certainembodiments, the subject has cancer cells that are PD-L1 negative. Insome embodiments, the subject never smoked. In certain embodiments, thesubject formerly smoked. In one embodiment, the subject currentlysmokes. In certain embodiments, the SCLC comprises a small cellcarcinoma. In certain embodiments, the SCLC comprises a combined smallcell carcinoma.

In some embodiments, the present methods comprise administering aneffective amount of an anti-PD-1 antibody or administering an effectiveamount of an anti-PD-L1 antibody and an effective amount of ananti-CTLA-4 antibody. An effective amount of an anti-PD-1 antibodyand/or an anti-CTLA-4 antibody can be a flat dose or a weight baseddose.

In some embodiments, the disclosure includes a method of treating acancer or a subject afflicted with cancer comprising administering ananti-PD-1 antagonist in combination with an anti-CD30 antibody to treatcancer. An “anti-PD-1 antagonist” as referred herein includes anymolecule that inhibits interaction between PD-1 (receptor) and PD-L1(ligand) such that the signal pathway of PD-1/PD-L1 is blocked. In otherembodiments, an anti-PD-1 antagonist is a PD-1-Fc fusion protein. Incertain embodiments, an anti-PD-1 antagonist includes an anti-PD-1fusion protein, an antisense molecule, a small molecule, a ribozyme, ora nanobody that inhibits or prevents interaction between PD-1 and PD-L1.

In certain embodiments, the therapy of the present disclosure (e.g.,administration of an anti-PD-1 antibody or administration of ananti-PD-1 antibody and an anti-CTLA-4 antibody) effectively increasesthe duration of survival of the subject. In some embodiments, theanti-PD-1 antibody or the anti-PD-1 antibody and anti-CTLA-4 antibodycombination therapy of the present disclosure increases theprogression-free survival of the subject. In certain embodiments, theanti-PD-1 antibody or the anti-PD-1 antibody and anti-CTLA-4 antibodycombination therapy of the present disclosure increases theprogression-free survival of the subject in comparison tostandard-of-care therapies. In some embodiments, the anti-PD-1 antibodyand anti-CTLA-4 antibody combination therapy of the present disclosureincreases the progression-free survival of the subject in comparison toan anti-PD-1 antibody alone (i.e., anti-PD-1 antibody monotherapy). Insome embodiments, the anti-PD-1 antibody and anti-CTLA-4 antibodycombination therapy of the present disclosure increases theprogression-free survival of the subject in comparison to otheranti-PD-1 antibody combinations.

In some embodiments, after the administration of an anti-PD-1 antibodyor administration of an anti-PD-1 antibody and an anti-CTLA-4 antibody,the subject having a tumor derived from an SCLC can exhibit an overallsurvival of at least about 2 months, at least about 3 months, at leastabout 4 months, at least about 5 months, at least about 6 months, atleast about 7 months, at least about 8 months, at least about 9 months,at least about 10 months, at least about 11 months, at least about 12months, at least about 13 months, at least about 14 months at leastabout 15 months, at least about 16 months, at least about 17 months, atleast about 18 months, at least about 19 months, at least about 20months, at least about 21 months, at least about 22 months, at leastabout 23 months, at least about 2 years, at least about 3 years, atleast about 4 years, or at least about 5 years after the administration.

In other embodiments, after the administration of a therapy disclosedherein (e.g., an anti-PD-1 antibody therapy or an anti-PD-1 antibody andan anti-CTLA-4 antibody therapy), the duration of survival or theoverall survival of the subject is increased by at least about 1 month,at least about 2 months, at least about 3 months, at least about 4months, at least about 6 months, or at least about 1 year when comparedto another subject treated with only a standard-of-care therapy (e.g., aplatinum-based chemotherapy) or a different dosing schedule of thetherapy. For example, the duration of survival or the overall survivalof the subject treated with an anti-PD-1 antibody disclosed herein isincreased by at least about 5%, at least about 10%, at least about 15%,at least about 20%, at least about 25%, at least about 30%, at leastabout 40%, at least about 50% or at least about 75% when compared toanother subject treated with only a standard-of-care therapy (e.g., aplatinum-based chemotherapy) or a different dosing schedule of theanti-PD-1 antibody therapy.

In other embodiments, after the administration of a combination therapycomprising an anti-PD-1 antibody and an anti-CTLA-4 antibody, theduration of survival or the overall survival of the subject is increasedby at least about 1 month, at least about 2 months, at least about 3months, at least about 4 months, at least about 6 months, or at leastabout 1 year when compared to another subject treated with only astandard-of-care therapy (e.g., a platinum-based chemotherapy), ananti-PD1 antibody alone, or a different dosing schedule of thecombination therapy. For example, the duration of survival or theoverall survival of the subject treated with an anti-PD-1 antibody andan anti-CTLA-4 antibody combination therapy disclosed herein isincreased by at least about 5%, at least about 10%, at least about 15%,at least about 20%, at least about 25%, at least about 30%, at leastabout 40%, at least about 50% or at least about 75% when compared toanother subject treated with only a standard-of-care therapy (e.g., aplatinum-based chemotherapy), an anti-PD-1 antibody alone, or adifferent dosing schedule of the combination therapy.

In certain embodiments, the therapy of the present disclosureeffectively increases the duration of progression free survival of thesubject. In some embodiments, the subject exhibits a progression-freesurvival of at least about one month, at least about 2 months, at leastabout 3 months, at least about 4 months, at least about 5 months, atleast about 6 months, at least about 7 months, at least about 8 months,at least about 9 months, at least about 10 months, at least about 11months, at least about one year, at least about eighteen months, atleast about two years, at least about three years, at least about fouryears, or at least about five years.

In some embodiments, the anti-PD-1 and anti-CTLA-4 antibodies areformulated for intravenous administration. In certain embodiments, theanti-PD-1 and anti-CTLA-4 antibodies are administered sequentially. Inembodiments, the anti-PD-1 and anti-CTLA-4 antibodies are administeredwithin 30 minutes of each other. In one embodiment, the anti-PD-1antibody is administered before the anti-CTLA-4 antibody. In anotherembodiment, the anti-CTLA-4 antibody is administered before theanti-PD-1 antibody. In another embodiment, the anti-PD-1 antibody andthe anti-CTLA-4 antibody are administered concurrently in separatecompositions. In a further embodiment, the anti-PD-1 antibody and theanti-CTLA-4 antibody are admixed as a single composition for concurrentadministration.

In some embodiments, the anti-PD-1 antibody and anti-CTLA-4 antibody areadministered in a fixed dose.

In some embodiments, the PD-L1 status of a tumor in a subject ismeasured prior to administering any composition or utilizing any methoddisclosed herein. In one embodiment, the PD-L1 expression level of atumor is at least about 1%, at least about 2%, at least about 3%, atleast about 4%, at least about 5%, at least about 6%, at least about 7%,at least about 8%, at least about 9%, at least about 10%, at least about11%, at least about 12%, at least about 13%, at least about 14%, atleast about 15%, at least about 20%, or greater than at least about 20%.In another embodiment, the PD-L1 status of a tumor is at least about 1%.In other embodiments, the PD-L1 status of the subject is at least about5%. In a certain embodiment, the PD-L1 status of a tumor is at leastabout 10%.

In some embodiments, the median progression-free survival of a subjectwith a tumor that has ≥1% PD-L1 expression is at least about 1 week, atleast about 2 weeks, at least about 3 weeks, at least about 4 weeks, atleast about 1 month, at least about 2 months, at least about 3 months,at least about 4 months, at least about 5 months, at least about 6months, at least about 7 months, at least about 8 months, at least about9 months, at least about 10 months, at least about 11 months, or atleast about 1 year longer than the median progression-free survival of asubject with a tumor with a <1% PD-L1 expression. In some embodiments,the progression-free survival of a subject with a tumor that has ≥1%PD-L1 expression is at least about 1 month, at least about 2 months, atleast about 3 months, at least about 4 months, at least about 5 months,at least about 6 months, at least about 7 months, at least about 8months, at least about 9 months, at least about 10 months, at leastabout 11 months, at least about 1 year, at least about eighteen months,at least about 2 years, at least about 3 years, at least about 4 years,or at least about 5 years.

In order to assess the PD-L1 expression, in one embodiment, a testtissue sample can be obtained from the patient who is in need of thetherapy. In another embodiment, the assessment of PD-L1 expression canbe achieved without obtaining a test tissue sample. In some embodiments,selecting a suitable patient includes (i) optionally providing a testtissue sample obtained from a patient with cancer of the tissue, thetest tissue sample comprising tumor cells and/or tumor-infiltratinginflammatory cells; and (ii) assessing the proportion of cells in thetest tissue sample that express PD-L1 on the surface of the cells basedon an assessment that the proportion of cells in the test tissue samplethat express PD-L1 on the cell surface is higher than a predeterminedthreshold level.

In any of the methods comprising the measurement of PD-L1 expression ina test tissue sample, however, it should be understood that the stepcomprising the provision of a test tissue sample obtained from a patientis an optional step. It should also be understood that in certainembodiments the “measuring” or “assessing” step to identify, ordetermine the number or proportion of, cells in the test tissue samplethat express PD-L1 on the cell surface is performed by a transformativemethod of assaying for PD-L1 expression, for example by performing areverse transcriptase-polymerase chain reaction (RT-PCR) assay or an IHCassay. In certain other embodiments, no transformative step is involvedand PD-L1 expression is assessed by, for example, reviewing a report oftest results from a laboratory. In certain embodiments, the steps of themethods up to, and including, assessing PD-L1 expression provides anintermediate result that may be provided to a physician or otherhealthcare provider for use in selecting a suitable candidate for theanti-PD-1 antibody or anti-PD-L1 antibody therapy. In certainembodiments, the steps that provide the intermediate result is performedby a medical practitioner or someone acting under the direction of amedical practitioner. In other embodiments, these steps are performed byan independent laboratory or by an independent person such as alaboratory technician.

In certain embodiments of any of the present methods, the proportion ofcells that express PD-L1 is assessed by performing an assay to determinethe presence of PD-L1 RNA. In further embodiments, the presence of PD-L1RNA is determined by RT-PCR, in situ hybridization or RNase protection.In other embodiments, the proportion of cells that express PD-L1 isassessed by performing an assay to determine the presence of PD-L1polypeptide. In further embodiments, the presence of PD-L1 polypeptideis determined by immunohistochemistry (IHC), enzyme-linked immunosorbentassay (ELISA), in vivo imaging, or flow cytometry. In some embodiments,PD-L1 expression is assayed by IHC. In other embodiments of all of thesemethods, cell surface expression of PD-L1 is assayed using, e.g., IHC orin vivo imaging.

Imaging techniques have provided important tools in cancer research andtreatment. Recent developments in molecular imaging systems, includingpositron emission tomography (PET), single-photon emission computedtomography (SPECT), fluorescence reflectance imaging (FRI),fluorescence-mediated tomography (FMT), bioluminescence imaging (BLI),laser-scanning confocal microscopy (LSCM) and multiphoton microscopy(MPM), will likely herald even greater use of these techniques in cancerresearch. Some of these molecular imaging systems allow clinicians tonot only see where a tumor is located in the body, but also to visualizethe expression and activity of specific molecules, cells, and biologicalprocesses that influence tumor behavior and/or responsiveness totherapeutic drugs (Condeelis and Weissleder, “In vivo imaging incancer,” Cold Spring Harb. Perspect. Biol. 2(12):a003848 (2010)).Antibody specificity, coupled with the sensitivity and resolution ofPET, makes immunoPET imaging particularly attractive for monitoring andassaying expression of antigens in tissue samples (McCabe and Wu,“Positive progress in immunoPET—not just a coincidence,” Cancer Biother.Radiopharm. 25(3):253-61 (2010); Olafsen et al., “ImmunoPET imaging ofB-cell lymphoma using 124I-anti-CD20 scFv dimers (diabodies),” ProteinEng. Des. Sel. 23(4):243-9 (2010)). In certain embodiments of any of thepresent methods, PD-L1 expression is assayed by immunoPET imaging. Incertain embodiments of any of the present methods, the proportion ofcells in a test tissue sample that express PD-L1 is assessed byperforming an assay to determine the presence of PD-L1 polypeptide onthe surface of cells in the test tissue sample. In certain embodiments,the test tissue sample is a FFPE tissue sample. In other embodiments,the presence of PD-L1 polypeptide is determined by IHC assay. In furtherembodiments, the IHC assay is performed using an automated process. Insome embodiments, the IHC assay is performed using an anti-PD-L1monoclonal antibody to bind to the PD-L1 polypeptide.

In one embodiment of the present methods, an automated IHC method isused to assay the expression of PD-L1 on the surface of cells in FFPEtissue specimens. This disclosure provides methods for detecting thepresence of human PD-L1 antigen in a test tissue sample, or quantifyingthe level of human PD-L1 antigen or the proportion of cells in thesample that express the antigen, which methods comprise contacting thetest sample, and a negative control sample, with a monoclonal antibodythat specifically binds to human PD-L1, under conditions that allow forformation of a complex between the antibody or portion thereof and humanPD-L1. In certain embodiments, the test and control tissue samples areFFPE samples. The formation of a complex is then detected, wherein adifference in complex formation between the test sample and the negativecontrol sample is indicative of the presence of human PD-L1 antigen inthe sample. Various methods are used to quantify PD-L1 expression.

In a particular embodiment, the automated IHC method comprises: (a)deparaffinizing and rehydrating mounted tissue sections in anautostainer; (b) retrieving antigen using a decloaking chamber and pH 6buffer, heated to 110° C. for 10 min; (c) setting up reagents on anautostainer; and (d) running the autostainer to include steps ofneutralizing endogenous peroxidase in the tissue specimen; blockingnon-specific protein-binding sites on the slides; incubating the slideswith primary antibody; incubating with a post-primary blocking agent;incubating with NovoLink Polymer; adding a chromogen substrate anddeveloping; and counterstaining with hematoxylin.

For assessing PD-L1 expression in tumor tissue samples, a pathologistexamines the number of membrane PD-L1⁺ tumor cells in each field under amicroscope and mentally estimates the percentage of cells that arepositive, then averages them to come to the final percentage. Thedifferent staining intensities are defined as 0/negative, 1+/weak,2+/moderate, and 3+/strong. Typically, percentage values are firstassigned to the 0 and 3+ buckets, and then the intermediate 1+ and 2+intensities are considered. For highly heterogeneous tissues, thespecimen is divided into zones, and each zone is scored separately andthen combined into a single set of percentage values. The percentages ofnegative and positive cells for the different staining intensities aredetermined from each area and a median value is given to each zone. Afinal percentage value is given to the tissue for each stainingintensity category: negative, 1+, 2+, and 3+. The sum of all stainingintensities needs to be 100%. In one embodiment, the threshold number ofcells that needs to be PD-L1 positive is at least about 100, at leastabout 125, at least about 150, at least about 175, or at least about 200cells. In certain embodiments, the threshold number or cells that needsto be PD-L1 positive is at least about 100 cells.

Staining is also assessed in tumor-infiltrating inflammatory cells suchas macrophages and lymphocytes. In most cases macrophages serve as aninternal positive control since staining is observed in a largeproportion of macrophages. While not required to stain with 3+intensity, an absence of staining of macrophages should be taken intoaccount to rule out any technical failure. Macrophages and lymphocytesare assessed for plasma membrane staining and only recorded for allsamples as being positive or negative for each cell category. Stainingis also characterized according to an outside/inside tumor immune celldesignation. “Inside” means the immune cell is within the tumor tissueand/or on the boundaries of the tumor region without being physicallyintercalated among the tumor cells. “Outside” means that there is nophysical association with the tumor, the immune cells being found in theperiphery associated with connective or any associated adjacent tissue.

In certain embodiments of these scoring methods, the samples are scoredby two pathologists operating independently, and the scores aresubsequently consolidated. In certain other embodiments, theidentification of positive and negative cells is scored usingappropriate software.

A histoscore is used as a more quantitative measure of the IHC data. Thehistoscore is calculated as follows:

Histoscore=[(% tumor×1 (low intensity))+(% tumor×2 (mediumintensity))+(% tumor×3 (high intensity)]

To determine the histoscore, the pathologist estimates the percentage ofstained cells in each intensity category within a specimen. Becauseexpression of most biomarkers is heterogeneous the histoscore is a truerrepresentation of the overall expression. The final histoscore range is0 (no expression) to 300 (maximum expression).

An alternative means of quantifying PD-L1 expression in a test tissuesample IHC is to determine the adjusted inflammation score (AIS) scoredefined as the density of inflammation multiplied by the percent PD-L1expression by tumor-infiltrating inflammatory cells (Taube et al.,“Colocalization of inflammatory response with B7-hl expression in humanmelanocytic lesions supports an adaptive resistance mechanism of immuneescape,” Sci. Transl. Med. 4(127):127ra37 (2012)).

The present methods can treat a tumor derived from an SCLC of anystages. There are at least nine stages used for SCLC: occult (hidden;TX, N0, and M0) stage, Stage 0 (carcinoma in situ; Tis, N0, and M0),Stage IA (T1a/T1b, N0, and M0), Stage IB (T2a, N0, M0), Stage IIA(T1a/T1b, N1, and M0; T2a, N1, and M0; or T2b, N0, and M0), Stage IIB(T2b, N1, and M0; or T3, NO, M0), Stage IIIA (T1-T3, N2, and M0; T3, N1,and M0; or T4, N0-N1, and M0), Stage IIIB (Any T, N3, M0; or T4, N2, andM0), and Stage IV (any T, any N, and M1a; or any T, any N, and M1b)(see, e.g.,http://www.cancer.org/cancer/lungcancer-smallcell/detailedguide/small-cell-lung-cancer-staging,last visited Jun. 2, 2016). In the occult stage, the cancer cannot beseen by imaging or bronchoscopy. In Stage 0, cancer cells are found inthe lining of the airways.

In one embodiment, the present methods treat a Stage I SCLC. Stage ISCLC is divided in Stage IA and IB. In Stage IA, the tumor is no largerthan 3 cm across, has not reached the membranes that surround the lungs,does not affect the main branches of the bronchi, and has not spread tolymph nodes or distant sites. In Stage IB, one or more of the followingis true: 1) the tumor is larger than 3 centimeters but not larger than 5centimeters; 2) the cancer has spread to the main bronchus but is notwithin 2 cm of the carina; 3) the tumor has grown into the visceralpleura (the membranes surrounding the lungs) and is not larger than 5cm; or 4) the tumor is partially clogging the airways (and is not largerthan 5 cm).

In another embodiment, the methods of the present disclosure treat aStage II SCLC. Stage II SCLC is divided into Stage IIA and IIB. In StageIIA, the cancer has either spread to the lymph nodes within the lungand/or around the area where the bronchus enters the lung (hilar lymphnodes). If the cancer has spread to the lymph nodes, then the cancer canonly have spread to the lymph nodes on the same side of the chest as thetumor. If the cancer has not spread to the lymph nodes, the cancer isStage IIA if the cancer has spread to the area where the bronchus entersthe lung and one or more of the following is true: 1) the tumor islarger than 3 centimeters and not larger than 5 centimeters; 2) thetumor has grown into a main bronchus, but is not within 2 cm of thecarina (and it is not larger than 5 cm); 3) the tumor has grown into thevisceral pleura (the membranes surrounding the lungs) and is not largerthan 5 cm; or 4) the tumor is partially clogging the airways (and is notlarger than 5 cm). The tumor is also considered Stage IIA if the cancerhas not spread to the lymph nodes and one or more of the following istrue: 1) the tumor is larger than 5 centimeters but not larger than 7centimeters; 2) the cancer has spread to a main bronchus and is at least2 centimeters away from where the carina; 3) the tumor has spread to thevisceral pleura (the membranes surrounding the lungs); or 4) the tumoris partially clogging the airways (and is between 5 and 7 cm across). Instage IIB, the cancer has either spread to the lymph nodes or not. Ifthe cancer has spread to the lymph nodes, then the cancer can only havespread to the lymph nodes on the same side of the chest as the tumor,the lymph nodes with cancer are within the lung or near the bronchus andone or more of the following is true: 1) the tumor is larger than 5centimeters but not larger than 7 centimeters; 2) the tumor has spreadto a main bronchus and is at least 2 centimeters away from where thecarina; 3) the tumor has spread into the visceral pleura (the membranessurrounding the lungs) and is between 5 and 7 cm across; or 4) thecancer is partially clogging the airways (and is between 5 and 7 cmacross). The tumor is also considered Stage IIB if the cancer has notspread to the lymph nodes and one or more of the following is true: 1)the tumor is larger than 7 centimeters; 2) the cancer has grown into thechest wall, the breathing muscle that separates the chest from theabdomen (diaphragm), the membranes surrounding the space between thelungs (mediastinal pleura), or membranes of the sac surrounding theheart (parietal pericardium); 3) the cancer invades a main bronchus andis closer than 2 cm (about ¾ inch) to the carina, but it does notinvolve the carina itself; 4) the cancer has grown into the airwaysenough to cause an entire lung to collapse or to cause pneumonia in theentire lung; or 5) two or more separate tumor nodules are present in thesame lobe of a lung.

In other embodiments, any methods of the present disclosure treat StageIII SCLC, including Stage IIIA and/or Stage IIIB. Stage IIIA is dividedinto 3 sections. These 3 sections are based on 1) the size of the tumor;2) where the tumor is found and 3) which (if any) lymph nodes havecancer. In the first type of Stage IIIA SCLC, the main tumor can be anysize, and it has not grown into the space between the lungs(mediastinum), the heart, the large blood vessels near the heart (suchas the aorta), the windpipe (trachea), the tube connecting the throat tothe stomach (esophagus), the backbone, or the carina, nor has it spreadto different lobes of the same lung. Further, the cancer has spread tolymph nodes around the carina (the point where the windpipe splits intothe left and right bronchi) or in the space between the lungs(mediastinum), and these lymph nodes are on the same side as the mainlung tumor, but the cancer has not spread to distant sites. In thesecond type of Stage IIIA SCLC, the cancer has spread to the lymph nodeson the same side of the chest as the tumor, and the lymph nodes with thecancer are within the lung or near the bronchus. Additionally: 1) thetumor is larger than 7 cm across; 2) the cancer has grown into the chestwall, the breathing muscle that separates the chest from the abdomen(diaphragm), the membranes surrounding the space between the lungs(mediastinal pleura), or membranes of the sac surrounding the heart(parietal pericardium); 3) the cancer invades a main bronchus and iscloser than 2 cm to the carina, but it does not involve the carinaitself; 4) two or more separate tumor nodules are present in the samelobe of a lung; and 5) the cancer has grown into the airways enough tocause an entire lung to collapse or to cause pneumonia in the entirelung. In the third type of Stage IIIA SCLC, the cancer may or may nothave spread to lymph nodes within the lung and/or around the area wherethe bronchus enters the lung (hilar lymph nodes) and one or more of thefollowing is true: 1) a tumor of any size has grown into the spacebetween the lungs (mediastinum), the heart, the large blood vessels nearthe heart (such as the aorta), the windpipe (trachea), the tubeconnecting the throat to the stomach (esophagus), the backbone, or thecarina; and/or 2) two or more separate tumor nodules are present indifferent lobes of the same lung.

Stage IIIB is divided into 2 sections depending on 1) the size of thetumor, 2) where the tumor is found, and 3) which lymph nodes havecancer. In the first type of Stage IIIB SCLC, the cancer can be of anysize; it may or may not have grown into nearby structures or causedpneumonia or lung collapse; and it has spread to lymph nodes near thecollarbone on either side, and/or has spread to hilar or mediastinallymph nodes on the side opposite the primary tumor. But, the cancer hasnot spread to distant sites. In the second type of Stage IIIB SCLC, thecancer has also spread to lymph nodes around the carina (the point wherethe windpipe splits into the left and right bronchi) or in the spacebetween the lungs (mediastinum). Affected lymph nodes are on the sameside as the main lung tumor. It has not spread to distant sites. Inaddition, one or more of the following is true: 1) a tumor of any sizehas grown into the space between the lungs (mediastinum), the heart, thelarge blood vessels near the heart (such as the aorta), the windpipe(trachea), the tube connecting the throat to the stomach (esophagus),the backbone, or the carina; and/or 2) two or more separate tumornodules are present in different lobes of the same lung

In some embodiments, the methods of the disclosure treat a Stage IVSCLC. Stage IV SCLC is divided into 2 types. In the first type of StageIV SCLC, the tumor may be any size and one or more of the following istrue: 1) there are one or more tumors in both lungs; 2) cancer is foundin the fluid around the lungs or heart. In the second type of Stave IVSCLC, the cancer can be any size and may or may not have grown intonearby structures or reached nearby lymph nodes, and it has spread todistant lymph nodes or to other organs such as the liver, bones, orbrain.

Anti-PD-1 Antibodies or Anti-PD-L1 Antibodies

Anti-PD-1 antibodies suitable for use in the disclosed methods areantibodies that bind to PD-1 with high specificity and affinity, blockthe binding of PD-L1, and inhibit the immunosuppressive effect of thePD-1 signaling pathway. In any of the therapeutic methods disclosedherein, an anti-PD-1 or anti-PD-L1 “antibody” includes anantigen-binding portion that binds to the PD-1 or PD-L1 receptor,respectively, and exhibits the functional properties similar to those ofwhole antibodies in inhibiting ligand binding and upregulating theimmune system. In certain embodiments, the anti-PD-1 antibody orantigen-binding portion thereof cross-competes with nivolumab forbinding to human PD-1. In other embodiments, the anti-PD-L1 antibody orantigen-binding fragment thereof competes for binding with BMS-936559,MPDL3280A, MEDI4736, or MSB0010718C for binding to human PD-L1.

In other embodiments, the anti-PD-1 antibody, or anti-PD-L1 antibody, orantigen-binding portions thereof is a chimeric, humanized, or humanmonoclonal antibody or a portion thereof. In certain embodiments fortreating a human subject, the antibody is a humanized antibody In otherembodiments for treating a human subject, the antibody is a humanantibody. Antibodies of an IgG1, IgG2, IgG3, or IgG4 isotype can beused.

In certain embodiments, the anti-PD-1 antibody, or anti-PD-L1 antibody,or antigen-binding portions thereof comprises a heavy chain constantregion which is of a human IgG1 or IgG4 isotype. In certain otherembodiments, the sequence of the IgG4 heavy chain constant region of theanti-PD-1 antibody, or anti-PD-L1 antibody, or antigen-binding portionsthereof contains an S228P mutation which replaces a serine residue inthe hinge region with the proline residue normally found at thecorresponding position in IgG1 isotype antibodies. This mutation, whichis present in nivolumab, prevents Fab arm exchange with endogenous IgG4antibodies, while retaining the low affinity for activating Fc receptorsassociated with wild-type IgG4 antibodies (Wang et al. In vitrocharacterization of the anti-PD-1 antibody nivolumab, BMS-936558, and invivo toxicology in non-human primates, Cancer Imm Res, 2(9):846-56(2014)). In yet other embodiments, the antibody comprises a light chainconstant region which is a human kappa or lambda constant region. Inother embodiments, the anti-PD-1 antibody, anti-PD-L1 antibody, orantigen-binding portions thereof is a monoclonal antibody or anantigen-binding portion thereof.

Human antibodies that bind specifically to PD-1 with high affinity havebeen disclosed in U.S. Pat. No. 8,008,449. Other anti-PD-1 monoclonalantibodies have been described in, for example, U.S. Pat. Nos.6,808,710, 7,488,802, 8,168,757 and 8,354,509, and PCT Publication No.WO 2012/145493. Each of the anti-PD-1 human antibodies disclosed in U.S.Pat. No. 8,008,449 has been demonstrated to exhibit one or more of thefollowing characteristics: (a) binds to human PD-1 with a K_(D) of1×10⁻⁷ M or less, as determined by surface plasmon resonance using aBiacore biosensor system; (b) does not substantially bind to human CD28,CTLA-4 or ICOS; (c) increases T-cell proliferation in a Mixed LymphocyteReaction (MLR) assay; (d) increases interferon-γ production in an MLRassay; (e) increases IL-2 secretion in an MLR assay; (f) binds to humanPD-1 and cynomolgus monkey PD-1; (g) inhibits the binding of PD-L1and/or PD-L2 to PD-1; (h) stimulates antigen-specific memory responses;(i) stimulates antibody responses; and (j) inhibits tumor cell growth invivo. Anti-PD-1 antibodies usable in the present disclosure includemonoclonal antibodies that bind specifically to human PD-1 and exhibitat least one, in some embodiments, at least five, of the precedingcharacteristics. In some embodiments, the anti-PD-1 antibody isnivolumab. In one embodiment, the anti-PD-1 antibody is pembrolizumab.

In one embodiment, the anti-PD-1 antibody is nivolumab. Nivolumab (alsoknown as “OPDIVO®”; formerly designated 5C4, BMS-936558, MDX-1106, orONO-4538) is a fully human IgG4 (S228P) PD-1 immune checkpoint inhibitorantibody that selectively prevents interaction with PD-1 ligands (PD-L1and PD-L2), thereby blocking the down-regulation of antitumor T-cellfunctions (U.S. Pat. No. 8,008,449; Wang et al. In vitrocharacterization of the anti-PD-1 antibody nivolumab, BMS-936558, and invivo toxicology in non-human primates, Cancer Imm Res, 2(9):846-56(2014)). In another embodiment, the anti-PD-1 antibody or fragmentthereof cross-competes with nivolumab. In other embodiments, theanti-PD-1 antibody or fragment thereof binds to the same epitope asnivolumab. In certain embodiments, the anti-PD-1 antibody has the sameCDRs as nivolumab.

In another embodiment, the anti-PD-1 antibody (or antigen-bindingportion thereof) cross-competes with pembrolizumab. In some embodiments,the anti-PD-1 antibody or fragment thereof binds to the same epitope aspembrolizumab. In certain embodiments, the anti-PD-1 antibody has thesame CDRs as pembrolizumab. In another embodiment, the anti-PD-1antibody is pembrolizumab. Pembrolizumab (also known as “KEYTRUDA®”,lambrolizumab, and MK-3475) is a humanized monoclonal IgG4 antibodydirected against human cell surface receptor PD-1 (programmed death-1 orprogrammed cell death-1). Pembrolizumab is described, for example, inU.S. Pat. Nos. 8,354,509 and 8,900,587; see alsohttp://www.cancer.gov/drugdictionary?cdrid=695789 (last accessed: Dec.14, 2014). Pembrolizumab has been approved by the FDA for the treatmentof relapsed or refractory melanoma.

In other embodiments, the anti-PD-1 antibody (or antigen-binding portionthereof) cross-competes with MEDI0680. In still other embodiments, theanti-PD-1 antibody or fragment thereof binds to the same epitope asMEDI0680. In certain embodiments, the anti-PD-1 antibody has the sameCDRs as MEDI0680. In other embodiments, the anti-PD-1 antibody isMEDI0680 (formerly AMP-514), which is a monoclonal antibody. MEDI0680 isdescribed, for example, in U.S. Pat. No. 8,609,089B2 or inhttp://www.cancer.gov/drugdictionary?cdrid=756047 (last accessed Dec.14, 2014).

In certain embodiments, the first antibody is an anti-PD-1 antagonist.One example of the anti-PD-1 antagonist is AMP-224, which is a B7-DC Fcfusion protein. AMP-224 is discussed in U.S. Publ. No. 2013/0017199 orinhttp://www.cancer.gov/publications/dictionaries/cancer-drug?cdrid=700595(last accessed Jul. 8, 2015).

In other embodiments, the anti-PD-1 antibody (or antigen-binding portionthereof) cross-competes with BGB-A317. In some embodiments, theanti-PD-1 antibody or fragment thereof binds the same epitope asBGB-A317. In certain embodiments, the anti-PD-1 antibody has the sameCDRs as BGB-A317. In certain embodiments, the anti-PD-1 antibody isBGB-A317, which is a humanized monoclonal antibody. BGB-A317 isdescribed in U.S. Publ. No. 2015/0079109.

In other embodiments, the anti-PD-1 antibody (or antigen-binding portionthereof) cross-competes with INCSHR1210 (SHR-1210). In some embodiments,the anti-PD-1 antibody binds to the same epitope as INCSHR1210(SHR-1210). In certain embodiments, the anti-PD-1 antibody has the sameCDRs as INCSHR1210 (SHR-1210). In certain embodiments, the anti-PD-1antibody is INCSHR1210 (SHR-1210), which is a human monoclonal antibody.INCSHR1210 (SHR-1210) is described in WO2015/085847.

In other embodiments, the anti-PD-1 antibody (or antigen-binding portionthereof) cross-competes with REGN-2810. In some embodiments, theanti-PD-1 antibody binds to the same epitope as REGN-2810. In certainembodiments, the anti-PD-1 antibody has the same CDRs as REGN-2810. Incertain embodiments, the anti-PD-1 antibody is REGN-2810, which is ahuman monoclonal antibody. REGN-2810 is described in WO2015/112800.

In other embodiments, the anti-PD-1 antibody (or antigen-binding portionthereof) cross-competes with PDR001. In some embodiments, the anti-PD-1antibody binds to the same epitope as PDR001. In certain embodiments,the anti-PD-1 antibody has the same CDRs as PDR001. In certainembodiments, the anti-PD-1 antibody is PDR001, which is a humanizedmonoclonal antibody. PDR001 is described in WO2015/112900.

In other embodiments, the anti-PD-1 antibody (or antigen-binding portionthereof) cross-competes with TSR-042 (ANB011). In some embodiments, theanti-PD-1 antibody binds to the same epitope as TSR-042 (ANB011). Incertain embodiments, the anti-PD-1 antibody has the same CDRs as TSR-042(ANB011). In certain embodiments, the anti-PD-1 antibody is TSR-042(ANB011), which is a humanized monoclonal antibody. TSR-042 (ANB011) isdescribed in WO2014/179664.

In other embodiments, the anti-PD-1 antibody (or antigen-binding portionthereof) cross-competes with STI-1110. In some embodiments, theanti-PD-1 antibody binds to the same epitope as STI-1110. In certainembodiments, the anti-PD-1 antibody has the same CDRs as STI-1110. Incertain embodiments, the anti-PD-1 antibody is STI-1110, which is ahuman monoclonal antibody. STI-1110 is described in WO2014/194302.

Anti-PD-1 antibodies usable in the disclosed methods also includeisolated antibodies that bind specifically to human PD-1 andcross-compete for binding to human PD-1 with nivolumab (see, e.g., U.S.Pat. Nos. 8,008,449 and 8,779,105; WO 2013/173223). The ability ofantibodies to cross-compete for binding to an antigen indicates thatthese antibodies bind to the same epitope region of the antigen andsterically hinder the binding of other cross-competing antibodies tothat particular epitope region. These cross-competing antibodies areexpected to have functional properties very similar those of nivolumabby virtue of their binding to the same epitope region of PD-1.Cross-competing antibodies can be readily identified based on theirability to cross-compete with nivolumab in standard PD-1 binding assayssuch as Biacore analysis, ELISA assays or flow cytometry (see, e.g., WO2013/173223).

In certain embodiments, the antibodies that cross-compete for binding tohuman PD-1 with, or bind to the same epitope region of human PD-1 as,nivolumab are monoclonal antibodies. For administration to humansubjects, these cross-competing antibodies are chimeric antibodies, orhumanized or human antibodies. Such chimeric, humanized or humanmonoclonal antibodies can be prepared and isolated by methods well knownin the art.

Anti-PD-1 antibodies usable in the methods of the disclosed disclosurealso include antigen-binding portions of the above antibodies. It hasbeen amply demonstrated that the antigen-binding function of an antibodycan be performed by fragments of a full-length antibody Examples ofbinding fragments encompassed within the term “antigen-binding portion”of an antibody include (i) a Fab fragment, a monovalent fragmentconsisting of the V_(L), V_(H), C_(L) and C_(H1) domains; (ii) a F(ab″)₂fragment, a bivalent fragment comprising two Fab fragments linked by adisulfide bridge at the hinge region; (iii) a Fd fragment consisting ofthe V_(H) and C_(H1) domains; (iv) a Fv fragment consisting of the V_(L)and V_(H) domains of a single arm of an antibody, or any combinationthereof.

Anti-PD-1 antibodies suitable for use in the disclosed compositions areantibodies that bind to PD-1 with high specificity and affinity, blockthe binding of PD-L1 and or PD-L2, and inhibit the immunosuppressiveeffect of the PD-1 signaling pathway. In any of the compositions ormethods disclosed herein, an anti-PD-1 “antibody” includes anantigen-binding portion or fragment that binds to the PD-1 receptor andexhibits the functional properties similar to those of whole antibodiesin inhibiting ligand binding and upregulating the immune system. Incertain embodiments, the anti-PD-1 antibody or antigen-binding portionthereof cross-competes with nivolumab for binding to human PD-1. Inother embodiments, the anti-PD-1 antibody or antigen-binding portionthereof is a chimeric, humanized, or human monoclonal antibody or aportion thereof. In certain embodiments, the antibody is a humanizedantibody. In other embodiments, the antibody is a human antibody.Antibodies of an IgG1, IgG2, IgG3, or IgG4 isotype can be used.

In certain embodiments, the anti-PD-1 antibody or antigen-bindingportion thereof comprises a heavy chain constant region which is of ahuman IgG1 or IgG4 isotype. In certain other embodiments, the sequenceof the IgG4 heavy chain constant region of the anti-PD-1 antibody orantigen-binding portion thereof contains an S228P mutation whichreplaces a serine residue in the hinge region with the proline residuenormally found at the corresponding position in IgG1 isotype antibodies.This mutation, which is present in nivolumab, prevents Fab arm exchangewith endogenous IgG4 antibodies, while retaining the low affinity foractivating Fc receptors associated with wild-type IgG4 antibodies (Wanget al. (2014)). In yet other embodiments, the antibody comprises a lightchain constant region which is a human kappa or lambda constant region.In other embodiments, the anti-PD-1 antibody or antigen-binding portionthereof is a monoclonal antibody or an antigen-binding portion thereof.In certain embodiments of any of the therapeutic methods describedherein comprising administration of an anti-PD-1 antibody, the anti-PD-1antibody is nivolumab. In other embodiments, the anti-PD-1 antibody ispembrolizumab. In other embodiments, the anti-PD-1 antibody is chosenfrom the human antibodies 17D8, 2D3, 4H1, 4A11, 7D3 and 5F4 described inU.S. Pat. No. 8,008,449. In still other embodiments, the anti-PD-1antibody is MEDI0680 (formerly AMP-514), AMP-224, or BGB-A31

In certain embodiments of any of the therapeutic methods describedherein comprising administration of an anti-PD-1 antibody, the anti-PD-1antibody is nivolumab. In other embodiments, the anti-PD-1 antibody ispembrolizumab. In other embodiments, the anti-PD-1 antibody is chosenfrom the human antibodies 17D8, 2D3, 4H1, 4A11, 7D3, and 5F4 describedin U.S. Pat. No. 8,008,449. In still other embodiments, the anti-PD-1antibody is MEDI0680 (formerly AMP-514), or AMP-224.

In certain embodiments, an anti-PD-1 antibody used in the methods can bereplaced with another PD-1 or anti-PD-L1 antagonist. For example,because an anti-PD-L1 antibody prevents interaction between PD-1 andPD-L1, thereby exerting similar effects to the signaling pathway ofPD-1, an anti-PD-L1 antibody can replace the use of an anti-PD-1antibody in the methods disclosed herein. Therefore, in one embodiment,the present disclosure is directed to a method for treating a subjectafflicted with a tumor derived from an SCLC (e.g., a recurrent SCLC)comprising administering to the subject a therapeutically effectiveamount an anti-PD-L1 antibody. In certain embodiments, the anti-PD-L1antibody is BMS-936559 (formerly 12A4 or MDX-1105) (see, e.g., U.S. Pat.No. 7,943,743; WO 2013/173223). In other embodiments, the anti-PD-L1antibody is MPDL3280A (also known as RG7446 and atezolizumab) (see,e.g., Herbst et al. (2013) J Clin Oncol 31(suppl):3000. Abstract; U.S.Pat. No. 8,217,149), MEDI4736 (also called Durvalumab; Khleif (2013) In:Proceedings from the European Cancer Congress 2013; Sep. 27-Oct. 1,2013; Amsterdam, The Netherlands. Abstract 802, See U.S. Pat. No.8,779,108 or US 2014/0356353, filed May 6, 2014), or MSB0010718C (alsocalled Avelumab; See US 2014/0341917). In other embodiments, theanti-PD-L1 antibody is CX-072 (also called CytomX; See WO2016/149201).

Combination Therapies with Anti-PD-1 or Anti-PD-L1 Antibodies

In certain embodiments, an anti-PD-1 antibody or anti-PD-L1 antibody isadministered in combination with one or more other anti-cancer agents.In certain embodiments, the one or more anti-cancer agents have beenadministered to the subject prior to the administration of the anti-PD-1or anti-PD-L1 antibodies or prior to the combination with the anti-PD-1or anti-PD-L1 antibodies. In certain embodiments, the one or moreanti-cancer agents were not effective in treating the cancer. In someembodiments, the other anti-cancer agent is any anti-cancer agentdescribed herein or known in the art. In certain embodiments, the otheranti-cancer agent is an anti-CTLA-4 antibody. In one embodiment, theother anti-cancer agent is a chemotherapy or a platinum-based doubletchemotherapy (PT-DC). In certain embodiments, the other anti-canceragent is an EGFR-targeted tyrosine kinase inhibitor (TKI). In oneembodiment, the other anti-cancer agent is an anti-VEGF antibody. Inother embodiments, the anti-cancer agent is a platinum agent (e.g.,cisplatin, carboplatin), a mitotic inhibitor (e.g., paclitaxel,albumin-bound paclitaxel, docetaxel, taxotere, docecad), a fluorinatedVinca alkaloid (e.g., vinflunine, javlor), vinorelbine, vinblastine,etoposide, or pemetrexed gemcitabin. In one embodiment, the otheranti-cancer agent is 5-flurouracil (5-FU). In certain embodiments, theother anti-cancer agent is any other anti-cancer agent known in the art.In some embodiments, two or more additional anti-cancer agents areadministered in combination with the anti-PD-1 or anti-PD-L1 antibody.In some embodiments, the PD-1 or PD-L1 antibody is combined withsurgical resection and/or radiation therapy.

Anti-CTLA-4 Antibodies

Anti-CTLA-4 antibodies of the instant disclosure bind to human CTLA-4 soas to disrupt the interaction of CTLA-4 with a human B7 receptor.Because the interaction of CTLA-4 with B7 transduces a signal leading toinactivation of T-cells bearing the CTLA-4 receptor, disruption of theinteraction effectively induces, enhances or prolongs the activation ofsuch T cells, thereby inducing, enhancing, or prolonging an immuneresponse.

Human antibodies that bind specifically to CTLA-4 with high affinityhave been disclosed in U.S. Pat. Nos. 6,984,720 and 7,605,238. Otheranti-CTLA-4 monoclonal antibodies have been described in, for example,U.S. Pat. Nos. 5,977,318, 6,051,227, 6,682,736, and 7,034,121. Theanti-CTLA-4 human antibodies disclosed in U.S. Pat. Nos. 6,984,720 and7,605,238 have been demonstrated to exhibit one or more of the followingcharacteristics: (a) binds specifically to human CTLA-4 with a bindingaffinity reflected by an equilibrium association constant (K_(a)) of atleast about 10⁷ M⁻¹, or about 10⁹ M⁻¹, or about 10¹⁰ M⁻¹ to 10¹¹ M⁻¹ orhigher, as determined by Biacore analysis; (b) a kinetic associationconstant (k_(a)) of at least about 10³, about 10⁴, or about 10⁵ m⁻¹ s⁻¹;(c) a kinetic disassociation constant (k_(d)) of at least about 10³,about 10⁴, or about 10⁵ m⁻¹ s⁻¹; and (d) inhibits the binding of CTLA-4to B7-1 (CD80) and B7-2 (CD86). Anti-CTLA-4 antibodies usable in thepresent disclosure include monoclonal antibodies that bind specificallyto human CTLA-4 and exhibit at least one, at least two or, in oneembodiment, at least three of the preceding characteristics. Anexemplary clinical anti-CTLA-4 antibody is the human monoclonal antibody10D1 (now known as ipilimumab and marketed as YERVOY®) as disclosed inU.S. Pat. No. 6,984,720. Ipilimumab is an anti-CTLA-4 antibody for usein the methods disclosed herein. Another anti-CTLA-4 antibody usable inthe present methods is tremelimumab.

An exemplary clinical anti-CTLA-4 antibody is the human monoclonalantibody 10D1 (now known as ipilimumab and marketed as YERVOY®) asdisclosed in U.S. Pat. No. 6,984,720. Ipilimumab is an anti-CTLA-4antibody for use in the methods disclosed herein. Ipilimumab is a fullyhuman, IgG1 monoclonal antibody that blocks the binding of CTLA-4 to itsB7 ligands, thereby stimulating T cell activation and improving overallsurvival (OS) in patients with advanced melanoma.

Another anti-CTLA-4 antibody useful for the present methods istremelimumab (also known as CP-675,206). Tremelimumab is human IgG2monoclonal anti-CTLA-4 antibody. Tremelimumab is described inWO/2012/122444, U.S. Publ. No. 2012/263677, or WO Publ. No. 2007/113648A2.

Anti-CTLA-4 antibodies usable in the disclosed methods also includeisolated antibodies that bind specifically to human PD-1 andcross-compete for binding to human CTLA-4 with ipilimumab ortremelimumab or bind to the same epitope region of human CTLA-4 asipilimumab or tremelimumab. In certain embodiments, the antibodies thatcross-compete for binding to human CTLA-4 with, or bind to the sameepitope region of human PD-1 as does ipilimumab or tremelimumab, areantibodies comprising a heavy chain of the human IgG1 isotype. Foradministration to human subjects, these cross-competing antibodies arechimeric antibodies, humanized antibodies, or human antibodies. Usableanti-CTLA-4 antibodies also include antigen-binding portions of theabove antibodies such as Fab, F(ab″)₂, Fd, or Fv fragments.

Ipilimumab (YERVOY®) is a fully human, IgG1 monoclonal antibody thatblocks the binding of CTLA-4 to its B7 ligands, thereby stimulating Tcell activation and improving overall survival (OS) in patients withadvanced melanoma (Hodi et al. (2010) N Engl J Med 363:711-23).Concurrent therapy with nivolumab and ipilimumab in a Phase 1 clinicaltrial produced rapid and deep tumor regression in a substantialproportion of patients with advanced melanoma, and was significantlymore effective than either antibody alone (Wolchok et al. (2013) N EnglJ Med 369(2):122-33; WO 2013/173223). However, it was hitherto not knownwhether this combination of immunoregulatory antibodies would besimilarly effective in other tumor types.

Combination of an Anti-PD-1 Antibody with an Anti-CTLA-4 Antibody forTreating SCLC

This disclosure provides combination therapy methods for treating atumor derived from an SCLC wherein an anti-PD-1 antibody is combinedwith another anti-cancer agent that is an antibody or an antigen-bindingportion thereof that binds specifically to CTLA-4 and inhibits CTLA-4activity. The combination of the anti-PD-1 antibody, nivolumab, and theanti-CTLA-4 antibody, ipilimumab, has been demonstrated herein (seeExample 1) to produce early, durable antitumor activity in SCLCpatients, particularly with specific dosing schedules. Accordingly, incertain embodiments, the anti-CTLA-4 antibody that is used incombination with the anti-PD-1 antibody is ipilimumab. In embodiments,the anti-CTLA-4 antibody is tremelimumab. In other embodiments, theanti-CTLA-4 antibody or antigen-binding portion thereof is an antibodyor portion thereof that cross-competes with ipilimumab for binding tohuman CTLA-4. In certain other embodiments, the anti-CTLA-4 antibody orantigen-binding portion thereof is a chimeric, humanized or humanmonoclonal antibody or a portion thereof. In yet other embodiments, theanti-CTLA-4 antibody or antigen-binding portion thereof comprises aheavy chain constant region that is of a human IgG1 or IgG4 isotype. Insome embodiments, the anti-CTLA-4 antibody comprises a heavy chainconstant region that is of a human IgG1 isotype.

Because of durability of the clinical effect previously demonstratedwith immunotherapy by inhibition of immune checkpoints (see, e.g., WO2013/173223), the combination treatment can include, in alternativeembodiments, a finite number of doses, e.g., about 1-10 doses, or caninvolve dosing at long intervals, e.g., once about every 3-6 months oronce about every 1-2 years or longer intervals.

In certain embodiments of the present methods, the anti-PD-1 antibody isnivolumab. In other embodiments, it is pembrolizumab. In yet otherembodiments, the anti-CTLA-4 antibody is ipilimumab. In furtherembodiments, the anti-CTLA-4 antibody is tremelimumab. Typically, theanti-PD-1 and anti-CTLA-4 antibodies are formulated for intravenousadministration. In certain embodiments, when the anti-PD-1 andanti-CTLA-4 antibodies are administered in combination, they areadministered within 30 minutes of each other. Either antibody can beadministered first, that is, in certain embodiments, the anti-PD-1antibody is administered before the anti-CTLA-4 antibody, whereas inother embodiments, the anti-CTLA-4 antibody is administered before theanti-PD-1 antibody. Typically, each antibody is administered byintravenous infusion over a period of 60 minutes. In certainembodiments, the anti-PD-1 and anti-CTLA-4 antibodies are administeredconcurrently, either admixed as a single composition in apharmaceutically acceptable formulation for concurrent administration,or concurrently as separate compositions with each antibody in apharmaceutically acceptable formulation.

In some embodiments, the anti-PD-1 antibody is administered at a dose of1 mg/kg once every three weeks, and the anti-CTLA-4 antibody isadministered at a dose of 3 mg/kg once every three weeks. In otherembodiments, the 1 mg/kg dose of the anti-PD-1 antibody and the 3 mg/kgdose of the anti-CTLA-4 antibody are administered one dose for each, twodoses for each, three doses for each, four doses for each, five dosesfor each, six doses for each, seven doses for each, eight doses foreach, nine doses for each, or ten doses for each. In furtherembodiments, the combination therapy of the anti-PD-1 antibody and theanti-CTLA-4 antibody is followed by a monotherapy of an anti-PD-1antibody, e.g., at a dose of 3 mg/kg once every two weeks.

In certain embodiments, the anti-PD-1 antibody or antigen-bindingportion thereof is administered at a subtherapeutic dose. In certainother embodiments, the anti-CTLA-4 antibody or antigen-binding portionthereof is administered at a subtherapeutic dose. In furtherembodiments, both the anti-PD-1 antibody or antigen-binding portionthereof and the anti-CTLA-4 antibody or antigen-binding portion thereofare each administered at a subtherapeutic dose.

Standard-of-Care Therapies for SCLC

Standard-of-care therapies for different types of cancer are well knownby persons of skill in the art. For example, the National ComprehensiveCancer Network (NCCN), an alliance of 21 major cancer centers in theUSA, publishes the NCCN Clinical Practice Guidelines in Oncology (NCCNGUIDELINES®) that provide detailed up-to-date information on thestandard-of-care treatments for a wide variety of cancers (see NCCNGUIDELINES® (2014), available at:http://www.nccn.org/professionals/physician_gls/f_guidelines.asp, lastaccessed Jun. 2, 2016).

Surgery, radiation therapy (RT), and chemotherapy are the threemodalities commonly used to treat SCLC patients. The most commonly usedinitial chemotherapy regimen is etoposide (TOPOSAR® or VEPESID®) pluscisplatin (PLATINOL®), known as EP. For people with extensive-stagesmall cell lung cancer, chemotherapy alone using the EP regimen is thestandard treatment. However, another regimen that may be used iscarboplatin (PARAPLATIN®) plus irinotecan (CAMPTOSAR®).

Although SCLC is highly sensitive to initial treatments, includingchemotherapy and/or radiotherapy, most patients ultimately die due torecurrence of the SCLC. Therefore, there is a particular unmet needamong patients who have recurrent SCLC as there is a lack of aneffective treatment after first line therapy.

Pharmaceutical Compositions and Dosages

Therapeutic agents of the present disclosure can be constituted in acomposition, e.g., a pharmaceutical composition containing an antibodyand a pharmaceutically acceptable carrier. As used herein, a“pharmaceutically acceptable carrier” includes any and all solvents,dispersion media, coatings, antibacterial and antifungal agents,isotonic and absorption delaying agents, and the like that arephysiologically compatible. In one embodiment, the carrier for acomposition containing an antibody is suitable for intravenous,intramuscular, subcutaneous, parenteral, spinal or epidermaladministration (e.g., by injection or infusion). A pharmaceuticalcomposition of the disclosure can include one or more pharmaceuticallyacceptable salts, anti-oxidant, aqueous and non-aqueous carriers, and/oradjuvants such as preservatives, wetting agents, emulsifying agents, anddispersing agents.

Dosage regimens are adjusted to provide the optimum desired response,e.g., a maximal therapeutic response and/or minimal adverse effects. Foradministration of an anti-PD-1 antibody, as a monotherapy or incombination with another anti-cancer agent (e.g., in combination with ananti-CTLA-4 antibody), the dosage can range from about 0.01 to about 20mg/kg, about 0.1 to about 10 mg/kg, about 0.1 to about 5 mg/kg, about 1to about 5 mg/kg, about 2 to about 5 mg/kg, about 7.5 to about 12.5mg/kg, or about 0.1 to about 30 mg/kg of the subject's body weight. Forexample, dosages can be about 0.1, about 0.3, about 1, about 2, about 3,about 5 or about 10 mg/kg body weight, or about 0.3, about 1, about 2,about 3, or about 5 mg/kg body weight. The dosing schedule is typicallydesigned to achieve exposures that result in sustained receptoroccupancy (RO) based on typical pharmacokinetic properties of anantibody An exemplary treatment regime entails administration about onceper week, once about every 2 weeks, once about every 3 weeks, once aboutevery 4 weeks, once about a month, once about every 3-6 months orlonger. In certain embodiments, an anti-PD-1 antibody such as nivolumabis administered to the subject once about every 2 weeks. In otherembodiments, the antibody is administered once about every 3 weeks. Thedosage and scheduling can change during a course of treatment. Forexample, a dosing schedule for anti-PD-1 monotherapy can compriseadministering the antibody: (i) about every 2 weeks in about 6-weekcycles; (ii) about every 4 weeks for about six dosages, then about everythree months; (iii) about every 3 weeks; (iv) about 3 mg/kg to about 10mg/kg once followed by about 1 mg/kg every about 2-3 weeks. Consideringthat an IgG4 antibody typically has a half-life of 2-3 weeks, a dosageregimen for an anti-PD-1 antibody of the disclosure comprises at leastabout 0.3 mg/kg to at least about 10 mg/kg body weight, at least about 1mg/kg to at least about 5 mg/kg body weight, or at least about 1 mg/kgto at least about 3 mg/kg body weight via intravenous administration,with the antibody being given every about 14-21 days in up to about6-week or about 12-week cycles until complete response or confirmedprogressive disease. In certain embodiments, an anti-PD-1 monotherapy isadministered at 3 mg/kg every 2 weeks until progressive disease orunacceptable toxicity. In some embodiments, the antibody treatment, orany combination treatment disclosed herein, is continued for at leastabout 1 month, at least about 3 months, at least about 6 months, atleast about 9 months, at least about 1 year, at least about 18 months,at least about 24 months, at least about 3 years, at least about 5years, or at least about 10 years.

When used in combinations with other cancer agents (e.g., in combinationwith an anti-CTLA-4 antibody), the dosage of an anti-PD-1 antibody canbe lowered compared to the monotherapy dose. Dosages of nivolumab thatare lower than the typical 3 mg/kg, but not less than 0.001 mg/kg, aresubtherapeutic dosages. The subtherapeutic doses of an anti-PD-1antibody used in the methods herein are higher than 0.001 mg/kg andlower than 3 mg/kg. In some embodiments, a subtherapeutic dose is about0.001 mg/kg to about 1 mg/kg, about 0.01 mg/kg to about 1 mg/kg, about0.1 mg/kg to about 1 mg/kg, or about 0.001 mg/kg to about 0.1 mg/kg bodyweight. In some embodiments, the subtherapeutic dose is at least about0.001 mg/kg, at least about 0.005 mg/kg, at least about 0.01 mg/kg, atleast about 0.05 mg/kg, at least about 0.1 mg/kg, at least about 0.5mg/kg, or at least about 1.0 mg/kg body weight. Receptor-occupancy datafrom 15 subjects who received 0.3 mg/kg to 10 mg/kg dosing withnivolumab indicate that PD-1 occupancy appears to be dose-independent inthis dose range. Across all doses, the mean occupancy rate was 85%(range, 70% to 97%), with a mean plateau occupancy of 72% (range, 59% to81%). In some embodiments, 0.3 mg/kg dosing can allow for sufficientexposure to lead to maximal biologic activity. Receptor-occupancy datafrom 15 subjects who received 0.3 mg/kg to 10 mg/kg dosing withnivolumab indicate that PD-1 occupancy appears to be dose-independent inthis dose range. Across all doses, the mean occupancy rate was 85%(range, 70% to 97%), with a mean plateau occupancy of 72% (range, 59% to81%) (Brahmer et al. (2010) J Clin Oncol 28:3167-75). Thus, 0.3 mg/kgdosing can allow for sufficient exposure to lead to maximal biologicactivity.

Although higher nivolumab monotherapy dosing up to about 10 mg/kg everytwo weeks has been achieved without reaching the maximum tolerated does(MTD), the significant toxicities reported in other trials of checkpointinhibitors plus anti-angiogenic therapy (see, e.g., Johnson et al.(2013) Cancer Immunol Res 1:373-77; Rini et al. (2011) Cancer117:758-67) support the selection of a nivolumab dose lower than 10mg/kg.

In certain embodiments, the dose of an anti-PD-1 antibody (or ananti-PD-L1 antibody) is a fixed dose in a pharmaceutical composition. Inother embodiments, the method of the present disclosure can be used witha flat dose (a dose given to a patient irrespective of the body weightof the patient). For example, a flat dose of a nivolumab can be about240 mg. For example, a flat dose of pembrolizumab can be about 200 mg.In embodiments, the anti-PD-1 antibody or antigen-binding portionthereof is administered at a dose of about 240 mg. In embodiments, theanti-PD-1 antibody or antigen-binding portion thereof is administered ata dose of about 360 mg. In embodiments, the anti-PD-1 antibody orantigen-binding portion thereof is administered at a dose of about 480mg. In one embodiment, 360 mg of the anti-PD-1 antibody or antigenbinding fragment is administered once every 3 weeks. In anotherembodiment, 480 mg of the anti-PD-1 antibody or antigen binding fragmentis administered once every 4 weeks.

Ipilimumab (YERVOY®) is approved for the treatment of melanoma at 3mg/kg given intravenously once every 3 weeks for 4 doses. Thus, in someembodiments, about 3 mg/kg is the highest dosage of ipilimumab used incombination with the anti-PD-1 antibody though, in certain embodiments,an anti-CTLA-4 antibody such as ipilimumab can be dosed within the rangeof about 0.3 to about 10 mg/kg, about 0.5 to about 10 mg/kg, about 0.5to about 5 mg/kg, or about 1 to about 5 mg/kg body weight about everytwo or three weeks when combined with nivolumab. In other embodiments,ipilimumab is administered on a different dosage schedule fromnivolumab. In some embodiments, ipilimumab is administered about everyweek, about every two weeks, about every three weeks, about every 4weeks, about every five weeks, about every six weeks, about every sevenweeks, about every eight weeks, about every nine weeks, about every tenweeks, about every eleven weeks, about every twelve weeks or about everyfifteen weeks. Dosages of ipilimumab that are lower than the typical 3mg/kg every 3 weeks, but not less than 0.001 mg/kg, are subtherapeuticdosages. The subtherapeutic doses of an anti-CTLA-4 antibody used in themethods herein are higher than 0.001 mg/kg and lower than 3 mg/kg. Insome embodiments, a subtherapeutic dose is about 0.001 mg/kg to about 1mg/kg, about 0.01 mg/kg to about 1 mg/kg, about 0.1 mg/kg to about 1mg/kg, or about 0.001 mg/kg to about 0.1 mg/kg body weight. In someembodiments, the subtherapeutic dose is at least about 0.001 mg/kg, atleast about 0.005 mg/kg, at least about 0.01 mg/kg, at least about 0.05mg/kg, at least about 0.1 mg/kg, at least about 0.5 mg/kg, or at leastabout 1.0 mg/kg body weight. It has been shown that combination dosingof nivolumab at 3 mg/kg and ipilimumab at 3 mg/kg exceeded the MTD in amelanoma population, whereas a combination of nivolumab at 1 mg/kg plusipilimumab at 3 mg/kg or nivolumab at 3 mg/kg plus ipilimumab at 1 mg/kgwas found to be tolerable in melanoma patients (Wolchok et al., N Engl JMed 369(2):122-33 (2013)). Accordingly, although nivolumab is toleratedup to 10 mg/kg given intravenously every 2 weeks, in certain embodimentsdoses of the anti-PD-1 antibody do not exceed about 3 mg/kg whencombined with ipilimumab. In certain embodiments, based on risk-benefitand PK-PD assessments, the dosage used comprises a combination ofnivolumab at about 1 mg/kg plus ipilimumab at about 3 mg/kg, nivolumabat about 3 mg/kg plus ipilimumab at about 1 mg/kg, or nivolumab at about3 mg/kg plus ipilimumab at about 3 mg/kg is used, each administered at adosing frequency of once about every 2-4 weeks, in certain embodiments,once about every 2 weeks or once about every 3 weeks. In certain otherembodiments, nivolumab is administered at a dosage of about 0.1 mg/kg,about 0.3 mg/kg, about 1 mg/kg, about 2 mg/kg, about 3 mg/kg, or about 5mg/kg in combination with ipilimumab administered at a dosage of about0.1 mg/kg, about 0.3 mg/kg, about 1 mg/kg, about 2 mg/kg, about 3 mg/kg,or about 5 mg/kg, once about every 2 weeks, once about every 3 weeks, oronce about every 4 weeks.

In certain embodiments, the combination of an anti-PD-1 antibody and ananti-CTLA-4 antibody is administered intravenously to the subject in aninduction phase about every 2 or 3 weeks for 1, 2, 3 or 4administrations. In certain embodiments, the combination of nivolumaband ipilimumab is administered intravenously in the induction phaseabout every 2 weeks or about every 3 weeks for about 4 administrations.The induction phase is followed by a maintenance phase during which onlythe anti-PD-1 antibody is administered to the subject at a dosage ofabout 0.1 mg/kg, about 0.3 mg/kg, about 1 mg/kg, about 2 mg/kg, about 3mg/kg, about 5 mg/kg, or about 10 mg/kg about every two or three weeksfor as long as the treatment proves efficacious or until unmanageabletoxicity or disease progression occurs. In certain embodiments,nivolumab is administered during the maintenance phase at a dose ofabout 3 mg/kg body about every 2 weeks.

The antibodies disclosed herein can be administered according to a“treatment cycle” or a “cycle” (which terms are used interchangeablyherein). As used herein, the term “cycle” refers to a course oftreatment that is repeated on a regular schedule with periods of rest inbetween. For example, treatment given for one week followed by threeweeks of rest is a treatment cycle. In one embodiment, the anti-PD-1antibody and/or anti-CTLA-4 antibody is administered for at least 1, 2,3, 4, 5, 6, 7, 8, 9, or 10 cycles. In a particular embodiment, theadministration of the anti-PD-1 antibody and the anti-CTLA-4 antibodywas repeated four times (4 cycles).

In certain embodiments, the anti-PD-1 antibody and the anti-CTLA-4antibody is formulated as a single composition, wherein the dose of theanti-PD1 antibody and the dose of the anti-CTLA-4 antibody are combinedat a ratio of 1:50, 1:40, 1:30, 1:20, 1:10. 1:5, 1:3, 1:1, 3:1, 5:1,10:1, 20:1, 30:1, 40:1, or 50:1. In other embodiments, the dose of theanti-CTLA-4 antibody is a fixed dose. In certain embodiments, the doseof the anti-CTLA-4 antibody is a flat dose, which is given to a patientirrespective of the body weight. In a specific embodiment, the flat doseof the anti-CTLA-4 antibody is about 80 mg.

For combination of nivolumab with other anti-cancer agents, these agentsare administered at their approved dosages. Treatment is continued aslong as clinical benefit is observed or until unacceptable toxicity ordisease progression occurs. Nevertheless, in certain embodiments, thedosages of these anti-cancer agents administered are significantly lowerthan the approved dosage, i.e., a subtherapeutic dosage, of the agent isadministered in combination with the anti-PD-1 antibody The anti-PD-1antibody can be administered at the dosage that has been shown toproduce the highest efficacy as monotherapy in clinical trials, e.g.,about 3 mg/kg of nivolumab administered once about every three weeks(Topalian et al., N Engl J Med 366:2443-54 (2012a); Topalian et al.,Curr Opin Immunol 24:207-12 (2012b)), or at a significantly lower dose,i.e., at a subtherapeutic dose. In certain embodiments, the anti-PD-1antibody is administered at about 3 mg/kg once about every three weeks.

Dosage and frequency vary depending on the half-life of the antibody inthe subject. In general, human antibodies show the longest half-life,followed by humanized antibodies, chimeric antibodies, and non-humanantibodies. The dosage and frequency of administration can varydepending on whether the treatment is prophylactic or therapeutic. Inprophylactic applications, a relatively low dosage is typicallyadministered at relatively infrequent intervals over a long period oftime. Some patients continue to receive treatment for the rest of theirlives. In therapeutic applications, a relatively high dosage atrelatively short intervals is sometimes required until progression ofthe disease is reduced or terminated, or until the patient shows partialor complete amelioration of symptoms of disease. Thereafter, the patientcan be administered a prophylactic regime.

Actual dosage levels of the active ingredient or ingredients in thepharmaceutical compositions of the present disclosure can be varied soas to obtain an amount of the active ingredient which is effective toachieve the desired therapeutic response for a particular patient,composition, and mode of administration, without being unduly toxic tothe patient. The selected dosage level will depend upon a variety ofpharmacokinetic factors including the activity of the particularcompositions of the present disclosure employed, the route ofadministration, the time of administration, the rate of excretion of theparticular compound being employed, the duration of the treatment, otherdrugs, compounds and/or materials used in combination with theparticular compositions employed, the age, sex, weight, condition,general health, and prior medical history of the patient being treated,and like factors well known in the medical arts. A composition of thepresent disclosure can be administered via one or more routes ofadministration using one or more of a variety of methods well known inthe art. As will be appreciated by the skilled artisan, the route and/ormode of administration will vary depending upon the desired results.

In one embodiment, a subject treated with an anti-PD-1 antibody and ananti-CTLA-4 antibody in combination can be further treated with ananti-PD-1 antibody monotherapy.

Because anti-PD-1 and anti-PD-L1 target the same signaling pathway andhave been shown in clinical trials to exhibit similar levels of efficacyin a variety of cancers, including renal cell carcinoma (see Brahmer etal. (2012) N Engl J Med 366:2455-65; Topalian et al. (2012a) N Engl JMed 366:2443-54; WO 2013/173223), an anti-PD-L1 antibody may besubstituted for the anti-PD-1 antibody in any of the therapeutic methodsdisclosed herein. In certain embodiments, the anti-PD-L1 antibody isBMS-936559 (formerly 12A4 or MDX-1105) (see, e.g., U.S. Pat. No.7,943,743; WO 2013/173223). In other embodiments, the anti-PD-L1antibody is MPDL3280A (also known as RG7446 and atezolizumab) (see,e.g., Herbst et al. (2013) J Clin Oncol 31(suppl):3000. Abstract; U.S.Pat. No. 8,217,149) or MEDI4736 (Khleif (2013) In: Proceedings from theEuropean Cancer Congress 2013; Sep. 27-Oct. 1, 2013; Amsterdam, TheNetherlands. Abstract 802). In certain embodiments, the antibodies thatcross-compete for binding to human PD-L1 with, or bind to the sameepitope region of human PD-L1 as the above-references PD-L1 antibodiesare monoclonal antibodies. For administration to human subjects, thesecross-competing antibodies can be chimeric antibodies, or can behumanized or human antibodies. Such chimeric, humanized or humanmonoclonal antibodies can be prepared and isolated by methods well knownin the art.

Kits

Also within the scope of the present disclosure are kits comprising ananti-PD-1 antibody and another anti-cancer agent for therapeutic uses.Kits typically include a label indicating the intended use of thecontents of the kit and instructions for use. The term label includesany writing, or recorded material supplied on or with the kit, or whichotherwise accompanies the kit. Accordingly, this disclosure provides akit for treating a subject afflicted with a tumor derived from an SCLC,the kit comprising: (a) an amount ranging from about 4 mg to about 500mg of a PD-1 antibody or an antigen-binding portion thereof; and (b)instructions for using the PD-1 antibody or an antigen-binding portionthereof in any method disclosed herein. This disclosure further providesa kit for treating a subject afflicted with a tumor derived from anSCLC, the kit comprising: (a) an amount ranging from about 4 mg to about500 mg of a PD-1 antibody or an antigen-binding portion thereof; (b) anamount ranging from about 4 mg to about 500 mg of a CTLA-4 antibody oran antigen-binding portion thereof; and (c) instructions for using thePD-1 antibody or an antigen-binding portion thereof and the CTLA-4antibody or an antigen-binding portion thereof in any method disclosedherein. In some embodiments, the kit contains the PD-1 antibody or anantigen-binding portion thereof and the CTLA-4 antibody or anantigen-binding portion thereof as separation compositions. In someembodiments, the kit contains the PD-1 antibody or an antigen-bindingportion thereof and the CTLA-4 antibody or an antigen-binding portionthereof as a single composition. In certain embodiments, the anti-PD-1,and the anti-CTLA-4 antibody can be co-packaged in unit dosage form. Incertain embodiments for treating human patients, the kit comprises ananti-human PD-1 antibody disclosed herein, e.g., nivolumab orpembrolizumab. In other embodiments, the kit comprises an anti-humanCTLA-4 antibody disclosed herein, e.g., ipilimumab or tremelimumab.

The present disclosure is further illustrated by the following examplethat should not be construed as further limiting. The contents of allreferences cited throughout this application are expressly incorporatedherein by reference.

Example 1 Example 1

Small-cell lung cancer (SCLC), which accounts for approximately 14% ofall lung cancers, is strongly associated with tobacco use and has highmutation rates without known oncogenic drivers. Most patients presentwith extensive-stage disease characterized by widespread metastases andpoor survival. Although 35% to 86% of patients respond to first-linechemotherapy, disease progresses rapidly, and outcomes with second-linetreatment are poor.

Standard first-line chemotherapy for SCLC is a platinum-etoposidedoublet, with topotecan as second-line therapy in the United States (US)and European Union (EU) and amrubicin as second-line therapy in Japan.Though response rates with topotecan are 23% and 9% forplatinum-sensitive and platinum-resistant/refractory patients,respectively, they are not durable.

Nivolumab, a fully human IgG4 programmed death 1 (PD-1)immune-checkpoint-inhibitor antibody, significantly improved overallsurvival and had a favorable safety profile compared with docetaxel intwo phase 3 studies of patients with non-SCLC (NSCLC) who progressedafter first-line platinum-based doublet chemotherapy, leading to itsapproval in the US for treatment of patients with metastatic NSCLC andin the EU for treatment of patients with locally advanced or metastaticsquamous NSCLC. Ipilimumab, a fully human IgG1 cytotoxic T-lymphocyteantigen 4 (CTLA-4) immune-checkpoint inhibitor, significantly improvedoverall survival in two phase 3 studies in patients with advancedmelanoma, leading to approval in the US and the EU for this indication.

Preclinical data indicate that the combination of PD-1 and CTLA-4receptor blockade may improve antitumor activity, and the combination ofnivolumab plus ipilimumab has demonstrated deep and durable responses inseveral tumor types. The combination of nivolumab plus ipilimumab isapproved in the US for treatment of advanced melanoma. Based on theefficacy of combination treatment in melanoma, a clinical trial wasdesigned as a phase ½ trial to investigate the activity and safety ofnivolumab as monotherapy or in combination with ipilimumab in severaladvanced or metastatic tumor types. The evaluation of nivolumabmonotherapy and the combination of nivolumab and ipilimumab in patientswith advanced or metastatic solid tumors for which no standard of carein advanced lines of treatment exists will potentially generate evidenceof antitumor activity as a basis for further clinical development inthese tumor types. Here, we report activity, safety, and biomarkeranalyses for the SCLC cohort.

Methods Study Design and Participants

This was an international phase ½, two-stage, open-label multi-armtrial. Patients with SCLC were enrolled at 23 sites in six countries(Finland, Germany, Italy, Spain, UK, and US). Eligible patients hadhistologically or cytologically-confirmed limited or extensive-stageSCLC, with progressive disease after at least one platinum-basedchemotherapy regimen. Patients with platinum-sensitive orplatinum-resistant disease (relapse ≥ or <90 days after, or during,chemotherapy, respectively) were eligible regardless of programmeddeath-ligand 1 (PD-L1) expression. Patients were ≥18 years of age, withan Eastern Cooperative Oncology Group (ECOG) performance status of 0 or1 (0 to 5 scale: 0, no symptoms; 1, mild; higher numbers, greatertumor-related disability) and had adequate organ function. Patients wererequired to have measurable disease per the Response Evaluation Criteriain Solid Tumors (RECIST), version 1.1, and baseline tumor biopsy orarchival tumor material available for biomarker analyses. Tumor materialwas acceptable from biopsies performed before the screening period ifthe biopsy was done up to 3 months prior to start of treatment and noother systemic cancer therapy was administered in that time. Baselinelaboratory tests required to assess eligibility included white bloodcell counts, neutrophils, platelets, haemoglobin, serum creatinine,alanine aminotransferase (ALT), aspartate aminotransferase (AST), totalbilirubin, albumin, lipase, and amylase. Key exclusion criteria includedactive brain or leptomeningeal metastases, a history of autoimmunedisease (except for vitiligo, type I diabetes mellitus, residualhypothyroidism due to autoimmune thyroiditis only requiring hormonereplacement, or conditions not expected to recur in the absence of anexternal trigger), the need for immunosuppressive doses of systemiccorticosteroids (>10 mg per day prednisone equivalents) 2 weeks prior tostudy drug administration, and prior treatment with antibodies thatmodulate T-cell function or checkpoint pathways. Patients were alsoexcluded if they tested positive for hepatitis B virus or humanimmunodeficiency virus, and had unresolved toxicities from prioranticancer therapies.

Patient selection was not based on estimated survival. Median survivalfor patients with relapsed SCLC has been reported as approximately3.5-12 months.

The study protocol was approved by an institutional review board orethics committee at each participating center. The study was conductedin accordance with the Declaration of Helsinki and Good ClinicalPractice guidelines, as defined by the International Conference onHarmonisation. Prior to performing any study-specific procedures,written informed consent was obtained from all patients.

Procedures

Considerations for the dosing in the combination cohorts were asfollows: the 1 mg/kg nivolumab plus 3 mg/kg ipilimumab regimen is theapproved dose for the treatment of advanced melanoma; the 3 mg/kgnivolumab plus 1 mg/kg ipilimumab regimen was chosen to maximize thenivolumab dose based on nivolumab exposure response data (1 mg/kg vs. 3mg/kg); and to ensure that nivolumab plus ipilimumab is tolerable inpatients with SCLC, an initial dose-escalating safety evaluation stepwas performed (starting with 1 mg/kg nivolumab plus 1 mg/kg ipilimumab).The safety of the 1 mg/kg nivolumab plus 3 mg/kg ipilimumab and the 3mg/kg nivolumab plus 1 mg/kg ipilimumab regimens have been previouslyassessed in studies of other tumor types.

Patients with SCLC were assigned to one of the following treatmentcohorts: nivolumab as monotherapy at 3 mg per kilogram of body weight(nivolumab-3) administered intravenously every 2 weeks or combinationtreatment of nivolumab plus ipilimumab administered intravenously every3 weeks for 4 cycles, at dose level 1 (nivolumab 1 mg/kg+ipilimumab 1mg/kg [nivolumab-1/ipilimumab-1]), dose level 2 (nivolumab 1mg/kg+ipilimumab 3 mg/kg [nivolumab-1/ipilimumab-3]), or dose level 2b(nivolumab 3 mg/kg+ipilimumab 1 mg/kg [nivolumab-3/ipilimumab-1]),followed by 3 mg/kg of nivolumab every 2 weeks. See FIG. 1. To ensurethat the planned combination regimens would be tolerable in patientswith SCLC, an initial dose-escalating safety evaluation for thecombination arms was conducted. The first dose cohort was level 1. Ifthis was deemed tolerable, then level 2 was initiated. If dose level 2was deemed not tolerable, dose level 2b was investigated. Once thehighest dose level for further investigation was confirmed in thedose-escalating safety evaluation phase, the combination arms continuedenrolling patients. Patients on active treatment needed to be followedup for at least 6 weeks after the start of study treatment beforetolerability of a dose level was determined based on prespecifiedtolerability assessment criteria. However, tolerability beyond 6 weekswas also taken into consideration. For combination treatment, nivolumabwas administered first (60-minute infusion), followed by ipilimumab(90-minute infusion), as per previous studies evaluating nivolumab plusipilimumab. Patients received open-label treatment until diseaseprogression or occurrence of unacceptable toxicity (FIG. 1). Treatmentbeyond RECIST, version 1.1-defined progression was permitted if thepatient was tolerating and benefiting from treatment, based oninvestigator assessment. Using an interactive voice response system,patients were enrolled in one of the four cohorts in a sequentialmanner, or assigned if more than one cohort was open for enrollment.Patients progressing on nivolumab-3 could cross over to combinationcohorts.

No dose reductions or modifications were permitted for nivolumab oripilimumab. Criteria for dose delays (which were required forprotocol-defined reasons) and treatment discontinuation are detailed inthe appendix.

Tumor assessments by radiographic imaging were done at baseline, every 6weeks for the first 24 weeks, and every 12 weeks thereafter untildisease progression (investigator-assessed per RECIST, version1.1-defined progression) or treatment discontinuation. Survival wasmonitored continuously while patients were on treatment and every 3months after treatment discontinuation. Safety was evaluated throughoutthe study (Table 1), and adverse events were graded according to theNational Cancer Institute's Common Terminology Criteria for AdverseEvents, version 4.0.

TABLE 1 On-study safety assessment schedules in nivolumab andnivolumab/ipilimumab cohorts* Nivolumab Nivolumab/ Nivolumab/IpilimumabMonotherapy Ipilimumab Week 13 Onward Day 1 Week 1 to Week 12 Day 1Safety Timing Week 1, 3, 5, Day 1 Week 13, 15, 17, assessmentConsiderations 7, 9, etc. Week 1, 4, 7, 10 19, 21, etc. Targeted 72hours prior X X (+day 4, week 2, 5) X physical to dosing examinationVital signs 72 hours prior X X (+day 4, week 2, 5) X and oxygen todosing saturation Physical Weight prior to X X (+day 4, week 2, 5) Xmeasurements dosing Adverse continuously continuously continuouslyevents assessment Review of continuously continuously continuouslyconcomitant medications Laboratory 72 hours prior X X (+day 4, week 2,5) X tests* to dosing Pregnancy 24 hours prior X X X test to dosing,(baseline and (baseline and every 3 (baseline and every 4 for WOCBPevery 4 weeks) weeks) only weeks) *Complete blood count withdifferential, liver function tests, blood urea nitrogen or serum urealevel, creatinine, albumin, calcium, magnesium, sodium, potassium,chloride, lactic acid dehydrogenase, glucose, amylase, lipase, andthyroid stimulating hormone. WOCBP = women of child-bearing potential. X= assessment to be performed.

Tumor PD-L1 protein expression was assessed retrospectively inpretreatment (archival or fresh) tumor biopsy specimens with the use ofa validated, automated immunohistochemical assay (Dako North America,Carpinteria, Calif., USA) using a rabbit antihuman PD-L1 antibody (clone28-8, Epitomics Inc, Burlingame, Calif., USA). Tumor PD-L1 expressionwas categorized as positive when staining of tumor-cell membranes (atany intensity) was observed at prespecified expression levels of ≥1% or≥5% of tumor cells in a section that included ≥100 evaluable tumorcells. In the initial study protocol, analysis of the specimen was notrequired in advance of patient randomization; the protocol was laterrevised and this was made a requirement via a study amendment (for allcohorts of the study).

Outcomes

The primary endpoint of this study was the proportion of patients with aconfirmed objective response (defined as the number of patients with abest overall response of complete response or partial response [as perinvestigator-assessed RECIST, version 1.1 criteria] divided by thenumber of assigned patients). The objective response rate was theprimary endpoint as the trial objective was to evaluate antitumoractivity of nivolumab monotherapy or in combination with ipilimumab.

The secondary endpoints included overall survival, progression-freesurvival, duration of response, and the rate of treatment-relatedadverse events leading to treatment discontinuation. Overall survivalwas defined as the time between the date of treatment assignment and thedate of death due to any cause. Progression-free survival was defined asthe time from treatment assignment to the date of the first documentedtumor progression, as determined by the investigator (per RECIST,version 1.1), or death due to any cause, whichever occurred first.Duration of response was defined as the time from a best overallresponse of partial or complete response until the date progressivedisease was documented (using RECIST version 1.1) or death due to anycause. The correlation between PD-L1 expression by tumor cells andantitumor activity was a prespecified exploratory endpoint.

All activity analyses were performed on the basis of the originaltreatment assignment, not by crossover status.

Statistical Analysis

In parallel to the safety evaluation phase for the combination arms (asdescribed in Procedures), enrollment of patients followed a Simontwo-stage design (see Simon R., Control Clin. Trials 10:1-10 (1989)).This design was used to test whether nivolumab and/or the combination ofnivolumab and ipilimumab yields an objective response rate that is ofclinical interest in each of the tumor types; it also limits theexpected number of patients who receive treatment when the true responserate is not of clinical value. The two-stage test was conductedindependently in each cohort.

For each cohort, the Simon design requires 18 treated patients for thefirst stage and calls for termination of a cohort at stage 1 if there isless than one responder among the 18 treated patients within the cohort.Otherwise, if two or more responders are identified in up to 18 patientsin a cohort, additional patients will be assigned, to a total of 40treated patients in that cohort. The treatment will be considered ofclinical interest if, at the end of the second stage, there are eight orgreater responders among 40 treated patients in any single cohort.

Only treatment arms that met an objective response rate thresholdproceeded from stage 1 to stage 2. Enrollment in stage 2 in a giventreatment arm could continue even if the other treatment arm was stillin stage 1.

For stage 2, upon completion of enrollment for the initial 40 patients,additional patients were assigned into the nivolumab monotherapy arm andthe combination arms up to a total of 100 patients (including thoseassigned in stage 1) in each treatment arm. When nivolumab monotherapyor nivolumab-1/ipilimumab-3 proceeded to stage 2, assessment of doselevel 2b in stage 2 (nivolumab-3/ipilimumab-1) was initiated.

All analyses included treated patients who were enrolled at least 90days prior to database lock. All activity analyses were performed on thebasis of the original treatment assignment, not by crossover status.

Objective response rates were summarized by a binomial response rate andcorresponding two-sided 95% exact confidence interval (CI) using theClopper-Pearson method. Progression-free survival and overall survivalwere summarized descriptively using Kaplan-Meier methodology; medianvalues were estimated with two-sided 95% CIs, calculated using theBrookmeyer-Crowley method. Only treatment cohorts with more than sixpatients are represented in Kaplan-Meier plots. Patient with less than12 weeks follow-up were excluded from Kaplan-Meier plots.Progression-free survival and overall survival rates were also estimatedwith two-sided 95% CIs, calculated using the Greenwood formula. Durationof response was summarized using the Kaplan-Meier product-limit method.For PD-L1 biomarker analysis, best overall response was summarized foreach cohort by baseline PD-L1 expression and objective response rates,with exact 95% CIs computed using the Clopper-Pearson method. Allstatistical analyses were performed using SAS software (version 9.02).

Results

We enrolled and treated 216 patients with SCLC: 98 patients in thenivolumab 3 mg/kg cohort, three patients in the nivolumab 1 mg/kg plusipilimumab 1 mg/kg cohort, 61 patients in the nivolumab 1 mg/kg plusipilimumab 3 mg/kg cohort, and 54 in the nivolumab 3 mg/kg plusipilimumab 1 mg/kg cohort (FIG. 1). Three patients in the nivolumab 3mg/kg group, two patients in the nivolumab 1 mg/kg plus ipilimumab 3mg/kg group, and four patients in the nivolumab 3 mg/kg plus ipilimumab1 mg/kg group did not receive first-line platinum therapy and did notmeet eligibility criteria, but received treatment. None of the threepatients in the nivolumab 1 mg/kg plus ipilimumab 1 mg/kg cohortpermanently discontinued due to treatment-related adverse events withinthe first 6 weeks, allowing for enrollment in the other two combinationcohorts: nivolumab 1 mg/kg plus ipilimumab 3 mg/kg, and nivolumab 3mg/kg plus ipilimumab 1 mg/kg. At database lock, all patients had atleast 12 weeks of follow-up; median follow-up for patients continuing inthe study (including those who had died or discontinued treatment) was198.5 days (IQR 163.0-464.0) in the nivolumab 3 mg/kg cohort, 302 days(IQR not calculable) in the nivolumab 1 mg/kg plus ipilimumab 1 mg/kgcohort, 361.0 days (273.0-470.0) in the nivolumab 1 mg/kg plusipilimumab 3 mg/kg cohort, and 260.5 days (248.0-288.0) in the nivolumab3 mg/kg plus ipilimumab 1 mg/kg cohort (Table 2). Median follow-up forthe overall survival data is shown in the appendix (Table 2). Baselinecharacteristics are shown in Table 3; roughly half of patients had beentreated with two or more previous regimens, and about a third hadplatinum-resistant disease.

TABLE 2 Treatment exposure and patient disposition Nivolumab-1/Nivolumab-3/ Nivolumab-3 Ipilimumab-3 Ipilimumab-1 (n = 98) (n = 61) (n= 54) Median number of infusions Nivolumab 3.5 (2.0- 3.0 (2.0- 2.0 (2.0-6.0) 14.0) 6.0) Ipilimumab NA 3.0 (2.0- 2.0 (2.0- 4.0) 4.0) Medianfollow-up, 198.5 361.0 260.5 days* (163.0- (273.0- (248.0- 464.0) 470.0)288.0) Median follow-up 338.5 501.0 400.0 for overall survival, days**(303.0- (413.0- (387.0- 604.0) 610.0) 428.0) Pts Continuing 21 (21%) 19(31%) 11 (20%) Treatment Pts not Continuing  77 (79%)† 42 (69%) 43 (80%)Treatment Progressive disease 57 (58%) 26 (43%) 36 (67%) AE related to 4(4%) 7 (11%) 4 (7%) study drug AE unrelated to 10 (10%) 5 (8%) 1 (2%)study drug Death 0 2 (3%) 0 Patient request/ 5 (5%) 1 (2%) 2 (4%)withdrew consent Other 1 (1%) 1 (2%) 0 Patients continuing 66 (67%) 48(79%) 44 (82%) to be followed‡ Deaths 48 (49%) 30 (49%) 25 (46%) Datapresented as n, n (%) or median (IQR) unless otherwise stated. Allpatients were enrolled at least 90 days prior to database lock. AE =adverse event. IQR = interquartile range NA = not applicable. *Patientscontinuing in the study at the time of the November 6, 2015 databaselock. **Patients continuing in the study at the time of the March 24,2016 database lock (n = 98, nivolumab-3; n = 61,nivolumab-1/ipilimumab-3; n = 55, nivolumab-3/ipilimumab-1). †Onepatient with disease progression and one patient who requested todiscontinue treatment had treatment-related adverse events thatcontributed to discontinuation from treatment in the nivolumab-3 cohort.‡Includes patients still on treatment and patients off treatmentcontinuing in the follow-up period.

TABLE 3 Baseline patient characteristics Nivolumab-1/ Nivolumab-1/Nivolumab-3/ Nivolumab-3 Ipilimumab-1 Iipilimumab-3 Ipilimumab-1Baseline Characteristics (n = 98) (N = 3) (n = 61) (n = 54) Median age(IQR), years  63 (57-68)  61 (52-65) 66 (58-71)  61 (56-65) Age ≥75 9(9%) 0 7 (11%) 0 Sex Male 61 (62%) 2 (67%) 35 (57%) 32 (59%) Female 37(38%) 26 (43%) 22 (41%) Race White 91 (93%) 2 (67%) 60 (98%) 52 (96%)Black/African 3 (3%) 1 (33%) 1 (2%) 0 American Other 4 (4%) 0 0 1 (2%)Not reported 0 0 0 1 (2%) Prior treatment regimens 1 40 (41%) 1 (33%) 32(52%) 23 (43%) 2-3 55 (56%) 2 (67%) 23 (38%) 28 (52%) >3 3 (3%) 0  6(10%) 3 (6%) First-line platinum-treated patients* Platinum sensitive 55(56%) 1 (33%) 25 (41%) 21 (39%) Platinum resistant 30 (31%) 0 23 (38%)21 (39%) Unknown 10 (10%) 2 (67%) 11(18%)  8 (15%) Smoking StatusCurrent or former 95 (97%)  3 (100%) 57 (93%) 48 (89%) smoker Neversmoked 3 (3%) 4 (7%) 5 (9%) Unknown 0 0 1 (2%) PD-L1 expression level‡≥1% 10 (14%) 1 (50%)  9 (24%)  5 (13%) <1% 59 (86%) 1 (50%) 28 (76%) 35(88%) ≥5% 4 (6%) 0 2 (5%) 1 (3%) <5% 65 (94%)  2 (100%) 35 (95%) 39(98%) Indeterminate/ 29 (30%) 1 (33%) 24 (39%) 14 (26%) not evaluable/missing Data presented as n (%) or median (IQR) unless otherwise stated.*Three patients in the nivolumab 3 mg/kg group, two patients in thenivolumab 1 mg/kg plus ipilimumab 3 mg/kg group, and four patients inthe nivolumab 3 mg/kg plus ipilimumab 1 mg/kg group did not receivefirst-line platinum therapy and did not meet eligibility criteria,although they were treated and included in the analysis. †Defined as apatient who relapsed <90 days after chemotherapy. ‡Percentage of PD-L1evaluable patients; may exceed 100% due to rounding.

Patients received a median of 3 to 5 infusions of nivolumab (IQR2.0-6.0) in the nivolumab 3 mg/kg cohort, 9.0 infusions of nivolumab(IQR not calculable) and 4.0 infusions of ipilimumab (IQR notcalculable) in the nivolumab 1 mg/kg plus ipilimumab 1 mg/kg cohort, 3.0infusions each of nivolumab (2.0-14.0) and ipilimumab (2.0-4.0) in thenivolumab 1 mg/kg plus ipilimumab 3 mg/kg cohort, and 2.0 infusions eachof nivolumab (2.0-6.0) and ipilimumab (2.0-4.0) in the nivolumab 3 mg/kgplus ipilimumab 1 mg/kg cohort. At the time of analysis, 77 (79%)patients had discontinued nivolumab 3 mg/kg, 42 (69%) had discontinuednivolumab 1 mg/kg plus ipilimumab 3 mg/kg, and 43 (80%) had discontinuednivolumab 3 mg/kg plus ipilimumab 1 mg/kg; the most common reason wasdisease progression (FIG. 1; Table 4). Two patients discontinuednivolumab 1 mg/kg plus ipilimumab 1 mg/kg (one due to diseaseprogression, and one due to adverse event not related to study drug).

TABLE 4 Treatment exposure and patient disposition. Nivolumab1/Nivolumab-3/ Nivolumab-3 Ipilimumab-3 Ipilimumab-1 (n = 98) (n = 61) (n= 54) Median number of infusions Nivolumab 3.5 (2.0-6.0) 3.0 (2.0-14.0)2.0 (2.0-6.0) Ipilimumab NA 3.0 (2.0-4.0) 2.0 (2.0-4.0) Median 198.5361.0 260.5 follow-up, days* (163.0-464.0) (273.0-470.0) (248.0-288.0)Median follow-up 338.5 501.0 400.0 for OS, days** (303.0-604.0)(413.0-610.0) (387.0-428.0) Patients 21 (21%) 19 (31%) 11 (20%)continuing treatment Patients not  77 (79%)† 42 (69%) 43 (80%)continuing treatment Progressive disease 57 (58%) 26 (43%) 36 (67%) AErelated 4 (4%)  7 (11%) 4 (7%) to study drug AE unrelated 10 (10%) 5(8%) 1 (2%) to study drug Death 0 2 (3%) 0 Patient request/ 5 (5%) 1(2%) 2 (4%) withdrew consent Other 1 (1%) 1 (2%) 0 Patients continuing66 (67%) 48 (79%) 44 (82%) to be followed‡ Deaths 48 (49%) 30 (49%) 25(46%) Data presented as n, n (%) or median (IQR) unless otherwisestated. All patients were enrolled at least 90 days prior to databaselock. AE = adverse event. IQR =interquartile range NA = not applicable.*Patients continuing in the study at the time of the November 6, 2015database lock. **Patients continuing in the study at the time of theMarch 24, 2016 database lock (n = 98, nivolumab-3; n = 61,nivolumab-1/ipilimumab-3; n = 55, nivolumab-3/ipilimumab-1). †Onepatient with disease progression and one patient who requested todiscontinue treatment had treatment-related adverse events thatcontributed to discontinuation from treatment in the nivolumab-3 cohort.‡Includes patients still on treatment and patients off treatmentcontinuing in the follow-up period.

By blinded independent central review, ten (10% [95% CI 5-18]) of 98patients achieved a confirmed objective response with nivolumab 3 mg/kg,14 (23% [13-36]) of 61 with nivolumab 1 mg/kg plus ipilimumab 3 mg/kg,and ten (19% [9-31]) of 54 with nivolumab 3 mg/kg plus ipilimumab 1mg/kg (Table 5; FIGS. 3A-3C). One (33%) of three patients receivingnivolumab 1 mg/kg plus ipilimumab 1 mg/kg achieved an objective response(complete response; data not shown). The predefined threshold that twoor more of 18 patients in a particular group must have confirmed partialor complete response before continued enrollment for that group in stage2 was met. In the nivolumab 3 mg/kg cohort, seven patients died beforedisease assessment, four patients discontinued early (one due totoxicity, three due to clinical progression), and one patient withdrewconsent before completing the protocol; in the nivolumab 1 mg/kg plusipilimumab 3 mg/kg cohort, five patients died before disease assessment,one patient discontinued early due to clinical progression, one patientwas not evaluable because the first assessment was not performed, andone patient withdrew consent for scans and follow-up visits; and in thenivolumab 3 mg/kg plus ipilimumab 1 mg/kg cohort, two patients diedbefore disease assessment, three patients discontinued early (two due toclinical progression and one due to toxicity), and a CT scan was notperformed on one patient. The median duration of response was notreached (95% CI 4.4—not reached) with nivolumab 3 mg/kg, 7.7 months(4.0—not reached) with nivolumab 1 mg/kg plus ipilimumab 3 mg/kg, and4.4 months (3.7—not reached) with nivolumab 3 mg/kg plus ipilimumab 1mg/kg. 16 patients had a duration of response longer than 6 months: sixpatients in the nivolumab 3 mg/kg group, one patient in the nivolumab 1mg/kg plus ipilimumab 1 mg/kg group, eight patients in the nivolumab 1mg/kg plus ipilimumab 3 mg/kg group, and one patient in the nivolumab 3mg/kg plus ipilimumab 1 mg/kg group (median 9.6 months [IQR 7.1-14.3]).Median time to response is shown in Table 5. At the time of databaselock, eight (80%) of ten responses in the nivolumab 3 mg/kg group, oneof three responses in the nivolumab 1 mg/kg plus ipilimumab 1 mg/kggroup, seven (50%) of 14 responses in the nivolumab 1 mg/kg plusipilimumab 3 mg/kg group, and seven (70%) of ten responses in thenivolumab 3 mg/kg plus ipilimumab 1 mg/kg group were ongoing. 30patients in the nivolumab 3 mg/kg cohort, 15 in the nivolumab 1 mg/kgplus ipilimumab 3 mg/kg cohort, and six in the nivolumab 3 mg/kg plusipilimumab 1 mg/kg cohort continued treatment beyond progression.

TABLE 5 Tumor Response. Nivolumab-1/ Nivolumab-3/ Nivolumab-3Ipilimumab-3 Ipilimumab-1 (n = 98) (n = 61) (n = 54) Objective response;10 (10%; 14 (23%; 10 (19%; 5-18) 13-36) 9-31) 95% CI Best overallresponse Complete response 0 1 (2%) 0 Partial response 10 (10%) 13 (21%)10 (19%) Stable disease 22 (22%) 13 (21%)  9 (17%) Progressive disease52 (53%) 23 (38%) 29 (54%) Unable to determine 12 (12%)  8 (13%)  6(11%) Not reported 2 (2%) 3 (5%) 0 Time to objective   2.0 (1.3-2.8)  2.1 (1.4-2.8)   1.4 (1.3-2.7) response (IQR), months Data are n (%)unless otherwise stated. All patients were enrolled at least 90 daysprior to database lock.

As of the database lock on Mar. 24, 2016, 60 (61%) of 98 patients in thenivolumab 3 mg/kg cohort, 36 (59%) of 61 in the nivolumab 1 mg/kg plusipilimumab 3 mg/kg, and 35 (64%) of 55 in the nivolumab 3 mg/kg plusipilimumab 1 mg/kg had died. Median overall survival was 4.4 months (95%CI 3.0-9.3) in the nivolumab 3 mg/kg cohort, 7.7 months (3.6-18.0) inthe nivolumab 1 mg/kg plus ipilimumab 3 mg/kg cohort, and 6.0 months(3.6-11.0) in the nivolumab 3 mg/kg plus ipilimumab 1 mg/kg cohort.1-year overall survival was 33% (95% CI 22-45) for the nivolumab 3 mg/kgcohort, 43% (30-56) for the nivolumab 1 mg/kg plus ipilimumab 3 mg/kgcohort, and 35% (22-48) for the nivolumab 3 mg/g plus ipilimumab 1 mg/kgcohort (FIG. 4A).

76 (78%) patients in the nivolumab 3 mg/kg cohort, 44 (72%) in thenivolumab 1 mg/kg plus ipilimumab 3 mg/kg cohort, and 42 (78%) in thenivolumab 3 mg/kg plus ipilimumab 1 mg/kg cohort had disease progressionor died; median progression-free survival was 1.4 months (95% CI1.4-1.9), 2.6 months (1.4-4.1), and 1.4 months (1.3-2.2), respectively.1-year progression-free survival was 11% (95% CI 5-19) in the nivolumab3 mg/kg cohort and 19% (9-32) for the nivolumab 1 mg/kg plus ipilimumab3 mg/kg cohort (FIG. 4B). The nivolumab 3 mg/kg plus ipilimumab 1 mg/kgcohort had not met the 1-year milestone for progression-free survival atthe time of database lock. Two (67%) of three patients in the nivolumab1 mg/kg plus ipilimumab 1 mg/kg cohort had died and one (33%) had aprogression event. Nine patients crossed over from the nivolumab 3 mg/kgcohort to the combination cohorts after progression (one to nivolumab 1mg/kg plus ipilimumab 3 mg/kg and eight to nivolumab 3 mg/kg plusipilimumab 1 mg/kg); eight of these patients had further diseaseprogression and one patient in the nivolumab 3 mg/kg plus ipilimumab 1mg/kg cohort withdrew consent and therefore response could not bedetermined.

PD-L1 expression was assessable in 148 (69%) of 216 patient samples, ofwhich 39 (27%) were provided as fresh biopsies and 109 (74%) werearchived specimens. 25 (17%) had 1% or greater PD-L1 expression, andseven (5%) had 5% or greater PD-L1 expression (Table 3). In apre-planned exploratory analysis of the nivolumab 3 mg/kg, nivolumab 1mg/kg plus ipilimumab 3 mg/kg, and nivolumab 3 mg/kg plus ipilimumab 1mg/kg cohorts, tumor responses occurred in patients irrespective ofPD-L1 expression (FIGS. 5A-5C, respectively).

Grade 3 or 4 treatment-related adverse events occurred in 13 (13%) of 98patients in the nivolumab 3 mg/kg cohort, 18 (30%) of 61 patients in thenivolumab 1 mg/kg plus ipilimumab 3 mg/kg cohort, and ten (19%) of 54patients in the nivolumab 3 mg/kg plus ipilimumab 1 mg/kg cohort (Table6); no patients in the nivolumab 1 mg/kg plus ipilimumab 1 mg/kg cohorthad a grade 3 or 4 treatment-related adverse event (data not shown). Themost commonly reported grade 3 or 4 treatment-related adverse eventswere increased lipase (none vs. 5 [8%] vs. none) and diarrhea (none vs.3 [5%] vs. 1 [2%]). Four (4%) patients in the nivolumab 3 mg/kg cohort,two (67%) in the nivolumab 1 mg/kg plus ipilimumab 1 mg/kg cohort, 18(30%) in the nivolumab 1 mg/kg plus ipilimumab 3 mg/kg cohort, and eight(15%) in the nivolumab 3 mg/kg plus ipilimumab 1 mg/kg cohort had dosedelays due to treatment-related adverse events. Excluding malignantneoplasm, the most frequent serious adverse events were dyspnoea,experienced by five (5%) patients in the nivolumab 3 mg/kg cohort, two(3%) in the nivolumab 1 mg/kg plus ipilimumab 3 mg/kg cohort, and four(7%) in the nivolumab 3 mg/kg plus ipilimumab 1 mg/kg cohort, anddiarrhea, experienced by two (2%) patients in the nivolumab 3 mg/kgcohort, four (7%) in the nivolumab 1 mg/kg plus ipilimumab 3 mg/kgcohort, and four (7%) patients in the nivolumab 3 mg/kg plus ipilimumab1 mg/kg cohort. 17 patients discontinued treatment because oftreatment-related adverse events: six (6%) patients in the nivolumab 3mg/kg cohort (one patient each with limbic encephalitis, hyperglycaemia,stomatitis, increased alanine aminotransferase, increased gammaglutamyltransferase, and pneumonitis), seven (11%) in the nivolumab 1mg/kg plus ipilimumab 3 mg/kg cohort (one patient each with colitis,myasthenia gravis, pneumonitis, and cardiomyopathy, and uveitis; onepatient with hypothyroidism, hyperglycaemia and increased alanineaminotransferase; one patient with diarrhea and renal failure), and four(7%) in the nivolumab 3 mg/kg plus ipilimumab 1 mg/kg cohort (onepatient each with colitis, pneumonitis, and peripheral neuropathy; onepatient with dyspnoea and pneumonitis; Table 6). Two patients whoreceived nivolumab 1 mg/kg plus ipilimumab 3 mg/kg died fromtreatment-related events of myasthenia gravis and worsening of renalfailure, respectively, and one patient who received nivolumab 3 mg/kgplus ipilitnumab 1 mg/kg died from treatment-related pneumonitis. Otherthan because of disease progression and study drug toxicity, thefollowing deaths were reported: in the nivolumab 3 mg/kg group, three(3%) due to unknown causes, one (1%) due to sepsis and multi-organfailure, and one (1%) due to respiratory insufficiency not related totreatment; in the nivolumab 1 mg/kg plus ipilimumab 3 mg/kg group, three(5%) due to unknown causes, one (2%) due to subdural hematoma unrelatedto study drug, one (2%) due to sedation, one (2%) due to hypovolemicseptic shock and septic shock from candidaemia, and one (2%) due toabdominal sepsis and secondary intravascular disseminated coagulation;and in the nivolumab 3 mg/kg plus ipilimumab 1 mg/kg group, three (6%)due to unknown causes, and one (2%) due to adverse events not related tostudy drug.

TABLE 6 Treatment-related adverse events Nivolumab-3Nivolumab-1/Ipilimumab-3 Nivolumab-3/Ipilimumab-1 (n = 98) (n = 61) (n =54) Grade Grade Grade Grade Grade Grade Grade Grade Grade 1-2 3 4 1-2 34 1-2 3 4 Any event 39 (40%) 9 (9%) 4 (4%) 30 (49%) 14 (23%) 4 (7%) 30(56%)  8 (15%) 2 (4%) Fatigue 10 (10%) 1 (1%) 0 16 (26%) 0 0 12 (22%) 00 Pruritus 11 (11%) 0 0 11 (18%) 1 (2%) 0 5 (9%) 0 0 Diarrhea 7 (7%) 0 010 (16%) 3 (5%) 0 8 (15%) 1 (2%) 0 Nausea 7 (7%) 0 0 6 (10%) 1 (2%) 0 4(7%) 0 0 Decreased 6 (6%) 0 0 4 (7%) 0 0 6 (11%) 0 0 appetitePneumonitis 2 (2%) 1 (1%) 0 1 (2%) 1 (2%) 0 2 (4%) 0 1 (2%) Vomiting 2(2%) 1 (1%) 0 2 (3%) 1 (2%) 0 5 (9%) 0 0 Hypo- 3 (3%) 0 0 9 (15%) 1 (2%)0 4 (7%) 0 0 thyroidism Increased 3 (3%) 0 0 3 (5%) 0 0 0 1 (2%) 0aspartate amino- transferase Hyper- 2 (2%) 0 0 7 (11%) 0 0 3 (6%) 0 0thyroidism Hyponatraemia 2 (2%) 0 0 0 1 (2%) 0 0 0 0 Increased 2 (2%) 1(1%) 0 2 (3%) 0 0 0 1 (2%) 0 alanine amino- transferase Increased 2 (2%)0 0 0 0 0 1 (2%) 1 (2%) 0 transaminases Rash 2 (2%) 0 0 10 (16%) 2 (3%)0 4 (7%) 0 0 Anaemia 1 (1%) 0 0 4 (7%) 0 0 3 (6%) 1 (2%) 0 Dyspnoea 1(1%) 0 0 0 1 (2%) 0 1 (2%) 2 (4%) 0 Rash, 1 (1%) 0 0 6 (10%) 2 (3%) 0 2(4%) 0 0 maculopapular Adrenal 0 0 0 1 (2%) 0 0 1 (2%) 1 (2%) 0insufficiency Aseptic 0 0 0 0 0 0 0 0 1 (2%) meningitis Cardio- 0 0 0 00 1 (2%) 0 0 0 myopathy Colitis 0 0 0 1 (2%) 1 (2%) 0 0 1 (2%) 0Decreased 0 0 0 0 1 (2%) 0 0 0 0 neutrophil count Drug-induced 0 1 (1%)0 0 0 0 0 0 0 liver injury Encephalitis 0 0 1 (1%) 1 (2%) 0 0 0 0 0Eyelid ptosis 0 0 0 0 1 (2%) 0 0 0 0 Haemorrhagic- 0 0 0 0 1 (2%) 0 0 00 gastritis Hyper- 0 0 1 (1%) 2 (3%) 0 1 (2%) 0 0 0 glycaemiaHypertrans- 0 0 0 0 1 (2%) 0 0 0 0 aminasaemia Hypoxia 0 1 (1%) 0 0 0 00 0 0 Ileus 0 0 0 0 0 0 0 1 (2%) 0 Increased 0 0 1 (1%) 3 (5%) 1 (2%) 02 (4%) 0 0 amylase Increased 0 0 1 (1%) 0 0 0 0 1 (2%) 0 gamma glutamyl-transferase Inc reased 0 0 0 2 (3%) 4 (7%) 1 (2%) 0 0 0 lipase Largeintestine 0 0 0 0 0 0 0 1 (2%) 0 perforation Myasthenia 0 0 0 0 1 (2%) 00 0 0 gravis Non-cardiac 0 1 (1%) 0 0 0 0 0 0 0 chest pain Pericardial 01 (1%) 0 0 0 0 0 0 0 effusion Peripheral 0 0 0 0 0 0 0 1 (2%) 0neuropathy Renal failure 0 0 0 0 0 1 (2%) 0 0 0 Stomatitis 0 1 (1%) 0 1(2%) 0 0 0 0 0 Thrombo- 0 0 0 0 0 0 3 (6%) 1 (2%) 0 cytopenia Tumorlysis 0 0 0 0 0 0 0 1 (2%) 0 syndrome Data presented as n (%). Thistable reports grade 1-2 treatment-related events in ≥10% of patients inany treatment cohort and all grade 3-4 events. Safety analyses includedall patients who were enrolled at least 90 days prior to database lock;patients with adverse events after crossover from nivolumab 3 mg/kg tocombination treatment are excluded. Some patients had more than oneadverse event. Two patients in the nivolumab 1 mg/kg plus ipilimumab 3mg/kg group died from myasthenia gravis and worsening of renal failure,respectively; both events were regarded to be treatment related. Onepatient in the nivolumab 3 mg/kg plus ipilimumab 1 mg/kg cohort diedfrom pneumonitis, regarded as treatment related.

Two patients had grade 2 limbic encephalitis: one in the nivolumab 3mg/kg cohort (reported as not treatment-related by investigator) and onein the nivolumab 1 mg/kg plus ipilimumab 3 mg/kg cohort (reported astreatment-related by investigator); both events resolved withimmunosuppressive treatment. One patient in the nivolumab 3 mg/kg cohorthad grade 4 limbic encephalitis (reported as treatment-related byinvestigator) that did not resolve with intravenous immunoglobulin andcorticosteroid treatment. Treatment-related pneumonitis occurred ineight patients and resolved in six of eight patients with treatment. Theoutcome was unknown for one patient, and one patient died.

One patient who crossed over from nivolumab 3 mg/kg to nivolumab 1 mg/kgplus ipilimumab 3 mg/kg had treatment-related grade 3 elevations inalanine aminotransferase levels. Five (8%) patients in the nivolumab 1mg/kg plus ipilimumab 3 mg/kg cohort had grade 3 or 4 asymptomaticlipase elevations without clinical signs of pancreatitis (Table 6).

Objective responses were observed in patients with one previous line oftherapy, and in patients with two or more previous therapies (Table 7).Median overall survival and progression-free survival were notsubstantially different for patients with one versus two or moreprevious treatments, with the possible exception of longerprogression-free survival in patients with one previous therapyreceiving nivolumab 1 mg/kg plus ipilimumab 3 mg/kg (FIGS. 6A-6D).

TABLE 7 Best overall tumor response by lines of therapy. One PriorTherapy Nivolumab-1/ Nivolumab-3/ Nivolumab-3 Ipilimumab-3 Ipilimumab-1(n =40) (n =32) (n =23) Objective Response Rate  4 (10%)  9 (28%)  5(22%) (95% CI)    (3-24)     (14-47)    (8-44) Best overall responseComplete response 0 1 (3%) 0 Partial response  4 (10%)  8 (25%)  5 (22%)Stable disease  8 (20%)  6 (19%)  3 (13%) Progressive disease 22 (55%)10 (31%) 12 (52%) Unable to determine  5 (13%)  6 (19%)  3 (13%) Notreported 1 (3%) 1 (3%) 0 Two or More Prior Therapies Nivolumab-1/Nivolumab-3/ Nivolumab-3 Ipilimumab-3 Ipilimumab-1 (n = 58) (n = 29) (n= 31) Objective Response Rate  6 (10%)  5 (17%)  5 (16%) (95% CI)   (4-21)    (6-36)    (6-34) Best overall response Complete response 00 0 Partial response  6 (10%)  5 (17%)  5 (16%) Stable disease 14 (24%) 7 (24%)  6 (19%) Progressive disease 30 (52%) 13 (45%) 17 (55%) Unableto determine  7 (12%) 2 (7%)  3 (10%) Not reported 1 (2%) 2 (7%) 0 Datapresented as n or n (%) unless otherwise stated. All patients wereenrolled at least 90 days prior to database lock.

In a post-hoc analysis in patients treated with a platinum agent as afirst-line treatment, objective responses were achieved in patients withboth platinum-sensitive and platinum-resistant disease (FIGS. 7A-7C;Table 8). Among patients with platinum-sensitive disease, two (4%) of 55in the nivolumab 3 mg/kg cohort and two (8%) of 25 in the nivolumab 1mg/kg plus ipilimumab 3 mg/kg cohort received subsequent platinum-basedcancer therapy. No patients with platinum-sensitive disease in thenivolumab 3 mg/kg plus ipilimumab 1 mg/kg arm received subsequentplatinum-based cancer therapy.

TABLE 8 Best overall tumor response by sensitivity to first-lineplatinum-based treatment. Platinum Sensitive* Nivolumab-1/ Nivolumab-3/Nivolumab-3 Ipilimumab-3 Ipilimumab-1 ( n = 55) (n = 25) (n = 21)Objective  6 (11%)  7 (28%)  4 (19%) Response Rate (95% CI)    (4-22)   (12-49)    (5-42) Best overall response Complete 0 0 0 responsePartial response  6 (11%) 7 (28%)  4 (19%) Stable disease 14 (25%) 7(28%)  5 (24%) Progressive 29 (53%) 8 (32%) 11 (52%) disease Unable to 5(9%) 3 (12%) 1 (5%) determine Not reported 1 (2%) 0 0 PlatinumResistant† Nivolumab-1/ Nivolumab-3/ Nivolumab-3 Ipilimumab-3Ipilimumab-1 (n = 30) (n = 23) (n = 21) Objective  3 (10%)  4 (17%)  2(10%) response rate (95%CI)    (2-27)    (5-39)    (1-30) Best overallresponse Complete 0 1 (4%) 0 response Partial response  3 (10%)  3 (13%) 2 (10%) Stable disease  5 (17%) 2 (9%) 1 (5%) Progressive 16 (53%) 10(44%) 13 (62%) disease Unable to  5 (17%)  5 (22%)  5 (24%) determineNot reported 1 (3%) 2 (9%) 0 Data presented as n (%) unless otherwisestated. All patients were enrolled at least 90 days prior to databaselock. For patients with known response to platinum-based therapy,platinum sensitivity was unknown for 29 patients as follows:nivolumab-3, n = 10; nivolumab-1/ipilimumab-3, n = 11;nivolumab-3/ipilimumab-1, n = 8. *Patient relapsed ≥90 days afterplatinum-based chemotherapy. †Patient failed to respond to, or relapsed<90 days after, platinum-based chemotherapy.

As of the database lock on Mar. 30, 2017, by blinded independent centralreview, 11% of patients in the nivolumab 3 mg/kg group and 23% ofpatients in the nivolumab 1 mg/kg plus ipilimumab 3 mg/kg group had anobjective response to treatment. See Table 9. Median time to responsewas 1.4 months (1.1-4.1 months, range) in the nivolumab 3 mg/kg groupand 2 months (1-4.1 months, range) in the nivolumab 3 mg/kg plusipilimumab 1 mg/kg group. Median duration of response was 17.9 months(2.8-34.6, range) in the nivolumab 3 mg/kg group and 14.2 months(1.5-26.5, range) in the nivolumab 3 mg/kg plus ipilimumab 1 mg/kggroup. 45% of responders in the nivolumab 3 mg/kg group and 36% ofresponders in the nivolumab 3 mg/kg plus ipilimumab 1 mg/kg group had anongoing response after 2 years.

TABLE 9 Summary of response. Nivolumab Nivolumab + (n = 98) Ipilimumab(n = 61) ORR, % (95% CI) 11 (6, 19)  23 (13, 36) Median time toresponse,  1.4 (1.1-4.1)  2.0 (1.0-4.1) mo (range) Median DOR, mo(range)   17.9 (2.8-34.6+)   14.2 (1.5-26.5+) Patients with ongoing 4536 responses at 2 yr^(a), % ^(a)Percentage of responders (nivolumab, n =11; nivolumab + ipilimumab, n = 14)

Tumor PD-L1 expression was examined in patients who had an objectiveresponse to treatment. See Table 10. Within the non-randomized cohort,PD-L1 expression was not evaluable in 43 patients (27%). Of thosepatients with quantifiable PD-L1 expression (159 patients), 82% had atumor expressing less than 1% PD-L1 and 18% had a tumor expressing atleast 1% PD-L1. See FIG. 9. For those patients a tumor expressing atleast 1% PD-L1, there was a 9% ORR in the nivolumab 3 mg/kg group and a10% ORR in the nivolumab 1 mg/kg plus ipilimumab group 3 mg/kg. Forpatients with a tumor expressing less than 1% PD-L1, there was a 14% ORRin the nivolumab 3 mg/kg group and a 32% ORR in the nivolumab 1 mg/kgplus ipilimumab 3 mg/kg group.

TABLE 10 Objective response rate (ORR) by tumor PD-L1 expression ORR, %(n/N) Nivolumab + PD-L1 Nivolumab Ipilimumab expression (n = 98) (n =61) >ivol  9 (1/11) 10 (1/10) <1% 14 (9/64)  32 (10/31)

Overall survival (OS) was improved by treatment with nivolumab incombination with ipilimumab. See FIG. 9. Median overall survival was 4.1months (95% CI 3.0-6.8) in the nivolumab 3 mg/kg group, and 7.8 months(3.6-14.2) in the nivolumab 3 mg/kg plus ipilimumab 1 mg/kg group.1-year overall survival was 27% for the nivolumab 3 mg/kg group, and 40%for the nivolumab 1 mg/kg plus ipilimumab 3 mg/kg group. 2-year overallsurvival was 14% for the nivolumab 3 mg/kg group, and 26% for thenivolumab 1 mg/kg plus ipilimumab 3 mg/kg group. Minimum follow-upsoccurred at 19.6 months (median=23.3 months) for patients in thenivolumab 3 mg/kg group, and at 20.2 months (median=28.6 months) forpatients in the nivolumab 3 mg/kg plus ipilimumab 1 mg/kg group.Follow-up was calculated as time from first dose to database lock. Insome instances, follow-ups were more frequent, e.g., for patients whodied prior to the Mar. 30, 2017 database lock.

Discussion

Our findings show that nivolumab monotherapy and nivolumab plusipilimumab provide clinically meaningful activity and an acceptablesafety profile for patients with limited or extensive-stage SCLC anddisease progression after at least one prior regimen. The prognosis forpatients with progression after prior treatment with platinum-basedchemotherapy is poor. Patients with advanced SCLC frequently respond tofirst-line therapy; however, recurrence is inevitable, and effectiveoptions at the time of progression and in patients withplatinum-resistant disease are limited. Patients with extensive-stageSCLC have a 2-year survival rate of less than 5%.

Our trial enrolled a heterogeneous patient population withplatinum-sensitive or platinum-resistant disease and a range of previouslines of therapy, making comparisons with other second-line trialsdifficult. Responses and stable disease were seen in all treatmentcohorts. Tumor regression followed both conventional and immune-relatedpatterns of response (prolonged reduction in tumor burden in thepresence of new lesions). Although the numbers of patients in subgroupswere small, preliminary analysis showed similar responses betweenplatinum-sensitive and platinum-resistant subgroups, and similaractivity in patients with one previous regimen and those with two ormore previous regimens. Across treatment groups, responses were durable.

One phase 2 study evaluated temozolomide in a similar population ofpatients with disease progression after one or two previous chemotherapyregimens. Although the proportions of patients achieving an objectiveresponse were similar to those shown in our study—11 (23%) of 48patients with platinum-sensitive disease and two (13%) of 16 patientswith platinum-refractory disease—the median duration of response totemozolomide was lower: 3.5 months (range 1.4-14.7) for all treatedpatients. Rova-T, a DLL3-targeted antibody-drug conjugate, showedantitumor activity and manageable toxicity in a phase 1 study ofpatients with SCLC and progression after one or two previous lines oftherapy. An objective response was achieved in seven (44%) of 16patients positive for the DLL3 biomarker treated at the maximumtolerated doses.

Limitations of our study include that the study cohorts were notrandomized, and the study was not powered for formal comparisons acrosscohorts. Baseline characteristics were generally similar across thecohorts, and although the combination treatment cohorts showed similarresponses, responses seemed to be deeper with the nivolumab 1 mg/kg plusipilimumab 3 mg/kg regimen. This dosing regimen has also been shown tobe efficacious in previously untreated melanoma.

The activity of nivolumab as monotherapy or combined with ipilimumab inpatients irrespective of platinum sensitivity or line of therapy is animportant aspect differentiating immune-checkpoint inhibitors fromtopotecan or amrubicin in SCLC. Response to topotecan depends onchemosensitivity, driven by tumor resistance mutations. By contrast, thegenomically unstable nature of SCLC2 might make it sensitive toimmune-checkpoint blockade via induction or restoration of a tumorantigen-driven immune response. Because few lymphocytes are observed inSCLC tumors, one hypothesis is that there is a greater need to targetthe lymphoid compartment with CTLA-4 inhibition in addition to PD-1inhibition to maximize the treatment effect.

Some studies have shown increased activity of PD-1 blockade in patientswith PD-L1-expressing NSCLC. However, data, including from this study,suggest that there is a lower prevalence of PD-L1 expression in SCLCversus NSCLC. A trial of pembrolizumab, a PD-1 immune-checkpointinhibitor, reported an initial response in four (25%) of 16 patients anddurable responses in patients with PD-L1-positive extensive-stage SCLC.In our study, objective responses were observed in patients irrespectiveof PD-L1 expression, including deep tumor responses in patients withPD-L1 tumor expression less than 1%. Whether PD-L1 expression ispredictive of benefit in SCLC must await analysis in a largerpopulation.

Although more than half of patients in this trial had received two ormore chemotherapy regimens, 1-year overall survival (33% for nivolumab 3mg/kg and 43% for nivolumab 1 mg/kg plus ipilimumab 3 mg/kg) wascomparable with or better than that reported in historical trials ofsecond-line topotecan or amrubicin. Consistent with other trials withimmune-checkpoint inhibitors across multiple solid tumors, and unliketrials of topotecan, findings from our study showed a flattening of theoverall survival curves for the nivolumab 3 mg/kg and nivolumab 1 mg/kgplus ipilimumab 3 mg/kg cohorts, suggesting a survival benefit in asubset of patients. However, because of the small numbers in this trial,it is difficult to determine when this occurs. Also consistent withfindings from previous randomized trials of immuno-oncology agents,there seems to be a greater effect of nivolumab or ipilimumab treatmenton overall survival than progression-free survival.

Adverse events were managed using established safety guidelines. Mosttoxic effects in the nivolumab 3 mg/kg and nivolumab 3 mg/kg plusipilimumab 1 mg/kg cohorts were mild to moderate, with only six (6%)patients in the nivolumab 3 mg/kg group and four (7%) in the nivolumab 3mg/kg plus ipilimumab 1 mg/kg group discontinuing because of toxicity.More treatment-related grade 3 or 4 adverse events occurred in thenivolumab 1 mg/kg plus ipilimumab 3 mg/kg cohort, with seven (11%)patients discontinuing because of toxicity. This regimen was usedeffectively and safely in a phase 3 trial in patients with melanoma,suggesting that this schedule is feasible in patients with SCLC. In allcohorts, fewer treatment-related toxic effects were reported whencompared with trials of topotecan or amrubicin.

Three patients had limbic encephalitis, and one patient receivingnivolumab 1 mg/kg plus ipilimumab 3 mg/kg died of treatment-relatedmyasthenia gravis. Autoimmune encephalitis and myasthenia gravis havebeen reported, albeit rarely, with both nivolumab and ipilimumab. Thefrequency of these events seems to be higher in patients with SCLCcompared with other malignant diseases, perhaps due to the tendency forparaneoplastic neurological syndromes associated with this disease.Treatment-related pneumonitis was reported in eight patients across allthe treatment cohorts and resulted in death in one patient receivingnivolumab 3 mg/kg plus ipilimumab 1 mg/kg. It is crucial to closelymonitor for immune-related adverse events or unmasking of previouslysubclinical autoimmune disease processes, with prompt implementation ofsafety guidelines for effective management.

On the basis of these encouraging phase ½ data, phase 3 studiescomparing nivolumab (240 mg intravenously every 2 weeks) as a flat doseor nivolumab plus ipilimumab (1 mg/kg nivolumab and 3 mg/kg ipilimumabintravenously every 3 weeks for two 42-day cycles followed by nivolumab[240 mg intravenously every 2 weeks]) versus placebo as maintenancetherapy (in patients without progression) after first-line chemotherapy(CheckMate 451, NCT02538666), and for nivolumab (240 mg every 2 weeksintravenously) versus single-agent chemotherapy as second-line therapy(CheckMate 331, NCT02481830) in SCLC were initiated and are currentlyongoing.

Example 2

The phase ½ clinical trial detailed in Example 1 has been expanded toinclude a randomized cohort to further evaluate nivolumab as monotherapyor in combination with ipilimumab in patients with SCLC whose diseaseprogressed after platinum-based therapy. Here, we report an interimdescriptive analysis of the SCLC randomized cohort.

Results

Patients with SCLC, having received 1 or 2 prior platinum-containingtreatment regimens, were assigned to one of the following treatmentgroups in the randomized cohort: nivolumab as monotherapy at 3 mg perkilogram of body weight administered intravenously every 2 weeks, orcombination treatment of nivolumab plus ipilimumab administeredintravenously every 3 weeks for 4 cycles, at nivolumab 1 mg/kg andipilimumab 3 mg/kg, followed by 3 mg/kg of nivolumab every 2 weeks. Atotal of 242 patients were randomized 3:2 into the nivolumab monotherapytreatment group (n=147) or the nivolumab and ipilimumab combinationtherapy treatment group (n=95). See FIG. 8. The primary endpoint of thisstudy was the proportion of patients with a confirmed objective response(defined as the number of patients with a best overall response ofcomplete response or partial response [as per investigator-assessedRECIST, version 1.1 criteria] divided by the number of assignedpatients). The objective response rate was the primary endpoint as thetrial objective was to evaluate antitumor activity of nivolumabmonotherapy or in combination with ipilimumab. Median follow-up for thenivolumab 3 mg/kg group was 10.8 months and was 11.2 months for thenivolumab 1 mg/kg plus ipilimumab 3 mg/kg group.

Baseline characteristics for patients in the randomized cohort are shownin Table 11; all patients in this cohort had been treated with 1 or 2previous platinum-containing regimens.

TABLE 11 Advanced SCLC baseline patient characteristics-randomizedcohort (n = 147) (n = 95) Median age, years (range) 63.0 (29ge, 65.0(41ge, Median age, 44 51 Male, % 59 63 Prior treatment regimens, % 67 671 33 33 2-r Platinum sensitivity, % 50 42 Sensitive 49 57 Resistant 1 1Unknown/not reported Smoking status, % 92 95 Current/former smoker 7 4Never-smoker 1 1 Unknown ECOG performance status, % 33 28 0 67 71 1 0 1Not reported

Complete response (CR) was achieved in 2 patients in the randomizedcohort (1 patient in the nivolumab 3 mg/kg group and 1 patient in thenivolumab 1 mg/kg plus ipilimumab 3 mg/kg group). In the randomizedcohort, an objective response rate (ORR) of 12% (95% CI) was achieved inthe nivolumab 3 mg/kg group, and an ORR of 21% (95% CI) was achieved inthe nivolumab 1 mg/kg plus ipilimumab 3 mg/kg group in the randomizedcohort; whereas, in the non-randomized cohort, an ORR of 11% (95% CI)was achieved in the nivolumab 3 mg/kg group, and an ORR of 23% (95% CI)was achieved in the nivolumab 1 mg/kg plus ipilimumab 3 mg/kg group. SeeFIG. 10; Table 9. CIs are as follows: nivolumab (randomized): 7, 18;nivolumab+ipilimumab (randomized): 13, 31; nivolumab (non-randomized):6, 19; nivolumab+ipilimumab (non-randomized): 13, 36.

The median time to response in the randomized cohort was comparable tothat in the non-randomized cohort, i.e., 1.5 months in the nivolumab 3mg/kg group, and 1.4 months in the nivolumab 1 mg/kg plus ipilimumab 3mg/kg group.

In the randomized cohort, a progression free survival (PFS) rate of 18%(95% CI) was achieved in the nivolumab 3 mg/kg group, and a PFS rate of30% (95% CI) was achieved in the nivolumab 1 mg/kg plus ipilimumab 3mg/kg group in the randomized cohort; whereas, in the non-randomizedcohort, a PFS rate of 27% (95% CI) was achieved in the nivolumab 3 mg/kggroup, and a PFS rate of 36% (95% CI) was achieved in the nivolumab 1mg/kg plus ipilimumab 3 mg/kg group. See FIG. 11.

Overall survival (OS) rate in the randomized cohort was 65% (95% CI) inthe nivolumab 3 mg/kg group, and 64% (95% CI) in the nivolumab 1 mg/kgplus ipilimumab 3 mg/kg group; whereas, in the non-randomized cohort, OSwas 59% (95% CI) in the nivolumab 3 mg/kg group, and 72% (95% CI) in thenivolumab 1 mg/kg plus ipilimumab 3 mg/kg group. See FIG. 12. Theminimum follow-up time was 12 weeks.

Activity and safety of nivolumab monotherapy and nivolumab+ipilimumabcombination therapy was also assessed in pooled cohorts, i.e., thenivolumab 3 mg/kg groups from both the randomized cohort and thenon-randomized cohort (245 patients combined in the “pooled” nivolumab 3mg/kg group), and the nivolumab 1 mg/kg+ipilimumab 3 mg/kg groups fromboth the randomized cohort and the non-randomized cohort (156 patientscombined in the “pooled” nivolumab 1 mg/kg ipilimumab 3 mg/kg group).

The ORR was 11% (95% CI: 8, 6) in the pooled nivolumab 3 mg/kg group,and 22% (95% CI: 16, 29) in the pooled nivolumab 1 mg/kg+ipilimumab 3mg/kg group. See Table 12.

TABLE 12 ORR in Advanced SCLC by treatment subgroups in pooled cohortsNivolumab + Nivolumab Ipilimumab ORR, 95% ORR, 95% n % CI n % CI Overallpopulation 245 11 8, 16 156 22 16, 29 Line of therapy 1 prior therapy137 12 7, 18 98 19 12, 29 2 or more prior 108 11 6, 19 58 26 15, 39therapies Platinum sensitivity (all treated patients) Platinum-sensitive133 13 8, 20 85 26 17, 36 Platinum-resistant 110 10 5, 17 65 15  8, 26

Treatment-related adverse events (TRAEs) were assessed in the poolednivolumab 3 mg/kg group, and the pooled nivolumab 1 mg/kg+ipilimumab 3mg/kg group. Although TRAEs occurred in 55% of patients in the poolednivolumab 3 mg/kg group and 73% of patients in the pooled nivolumab 1mg/kg+ipilimumab 3 mg/kg group, grade 3-4 TRAEs were only observed in12% of patients in the pooled nivolumab 3 mg/kg group and 37% ofpatients in the pooled nivolumab 1 mg/kg+ipilimumab 3 mg/kg group. Ofthose, 78% (31/40 in the pooled nivolumab+ipilimumab group) and 45%(5/11 in the pooled nivolumab group) resolved. Median time to resolutionranged from 1.8 weeks (gastrointestinal events) to 16.3 weeks (hepaticevents) in the pooled nivolumab+ipilimumab group, and from 3.4 weeks(pulmonary events) to not reached (renal and hepatic events) in thepooled nivolumab group.

TABLE 13 Safety summary in Advanced SCLC by treatment subgroups inpooled cohorts Nivolumab (n = 245) Nivolumab + Ipilimumab (n = 156) Anygrade, % Grade 3de, % Any grade, % Grade 3de, % Any TRAEs 55 12 73 37TRAEs leading to 3  2 13 10 discontinuation Select TRAEs by categorySkin 16 <1 36 6 Endocrine 8  0 21 3 Hepatic 6  2 12 6 Gastrointestinal 5 0 24 8 Hypersensitivity/infusion 5  0 1 0 reaction Pulmonary 3  2 4 3Renal 1 <1 1 0

There were a total of 5 treatment-related deaths, 4 occurring in thepooled nivolumab+ipilimumab group due myasthenia gravis, pneumonitis,seizures/encephalitis, and autoimmune hepatitis (n=1 each) and 1occurring in the pooled nivolumab group due to pneumonitis. A previouslyreported death in the nivolumab+ipilimumab group due to renal failurewas subsequently determined to not be related to treatment.

Discussion

With a longer follow-up of greater than 2 years, the observed survivaland tumor responses to treatment with nivolumab monotherapy or nivolumaband ipilimumab combination therapy remained durable. Efficacy in therandomized cohort was consistent with that observed in thenon-randomized cohort. The response to treatment was observed regardlessof platinum sensitivity, line of therapy, and/or PD-L1 status.

What is claimed is:
 1. An antibody or an antigen-binding portion thereofthat binds specifically to a Programmed Death-1 (PD-1) receptor andinhibits PD-1 activity (“anti-PD-1 antibody”) for use in a method oftreating a subject afflicted with a tumor derived from a small cell lungcancer (SCLC), wherein the subject is administered the anti-PD-1antibody in combination with an antibody or an antigen-binding portionthereof that binds specifically to Cytotoxic T-Lymphocyte Antigen-4(CTLA-4) and inhibits CTLA-4 activity (“anti-CTLA-4 antibody”).
 2. Theanti-PD-1 antibody for use according to claim 1, wherein the SCLCcomprises a small cell carcinoma, a combined small cell carcinoma, or arecurrent SCLC.
 3. The anti-PD-1 antibody for use according to claim 1or 2, wherein at least one, at least two, at least three, at least four,or at least five previous lines of therapy were administered to treatthe tumor.
 4. The anti-PD-1 antibody for use according to claim 3,wherein the previous line of therapy comprises a chemotherapy,optionally a platinum-based therapy that preferably comprises aplatinum-based antineoplastic selected from the group consisting ofcisplatin, carboplatin, oxaliplatin, nedaplatin, triplatin tetranitrate,phenanthriplatin, picoplatin, satraplatin, and any combination thereof.5. The anti-PD-1 antibody for use according to any one of claims 1 to 4,wherein (i) the anti-PD-1 antibody cross-competes with nivolumab forbinding to human PD-1; (ii) the anti-PD-1 antibody binds to the sameepitope as nivolumab; (iii) the anti-PD-1 antibody is nivolumab; or (iv)the anti-PD-1 antibody is pembrolizumab.
 6. The anti-PD-1 antibody foruse according to any one of claims 1 to 5, wherein (i) the anti-CTLA-4antibody cross-competes with ipilimumab for binding to human CTLA-4;(ii) the anti-CTLA-4 antibody is ipilimumab; or (iii) the anti-CTLA-4antibody is tremelimumab.
 7. The anti-PD-1 antibody for use according toany one of claims 1 to 6, wherein (i) the anti-PD-1 antibody isadministered at a dose ranging from at least about 0.1 mg/kg to at leastabout 10.0 mg/kg body weight once about every 1, 2, 3, or 4 weeks; (ii)the anti-PD-1 antibody is administered at a dose of about 1 mg/kg orabout 3 mg/kg body weight; (iii) the anti-PD-1 antibody is administeredat a flat dose, optionally at least about 200, at least about 220, atleast about 240, at least about 260, at least about 280, at least about300, at least about 320, at least about 340, at least about 360, atleast about 380, at least about 400, at least about 420, at least about440, at least about 460, at least about 480, at least about 500 or atleast about 550 mg, about once every 1, 2, 3 or 4 weeks; (iv) theanti-PD-1 antibody is administered once about every 2 weeks; or (v) theanti-PD-1 antibody is administered once about every 3 weeks.
 8. Theanti-PD-1 antibody for use according to any one of claims 1 to 7,wherein (i) the anti-CTLA-4 antibody is administered at a dose rangingfrom at least about 0.1 mg/kg to at least about 10.0 mg/kg body weightonce about every 1, 2, 3, or 4 weeks; (ii) the anti-CTLA-4 isadministered at a dose of about 1 mg/kg or about 3 mg/kg body weight;(iii) the anti-CTLA-4 antibody is administered once about every 2 weeks;or (iv) the anti-CTLA-4 antibody is administered once about every 3weeks.
 9. The anti-PD-1 antibody for use according to claim 8, wherein(i) the anti-PD-1 antibody is administered at a dose of about 3 mg/kgbody weight once about every 3 weeks and the anti-CTLA-4 antibody isadministered at a dose of about 1 mg/kg body weight once about every 3weeks; (ii) the anti-PD-1 antibody is administered at a dose of about 1mg/kg body weight once about every 3 weeks and the anti-CTLA-4 antibodyis administered at a dose of about 3 mg/kg body weight once about every3 weeks; or (iii) the anti-PD-1 antibody is administered at a dose ofabout 1 mg/kg body weight once about every 3 weeks and the anti-CTLA-4antibody is administered at a dose of about 1 mg/kg body weight onceabout every 3 weeks.
 10. The anti-PD-1 antibody for use according toclaim 9, wherein the administration of the anti-PD-1 antibody and theanti-CTLA-4 antibody was repeated four times (4 cycles).
 11. Theanti-PD-1 antibody for use according to any one of claims 1 to 10,wherein the subject is further treated with an anti-PD-1 antibodymonotherapy following the treatment in claims 1 to
 10. 12. The anti-PD-1antibody for use according to any one of claims 1 to 11, wherein thesubject exhibits progression-free survival of at least about one month,at least about 2 months, at least about 3 months, at least about 4months, at least about 5 months, at least about 6 months, at least about7 months, at least about 8 months, at least about 9 months, at leastabout 10 months, at least about 11 months, at least about one year, atleast about eighteen months, at least about two years, at least aboutthree years, at least about four years, or at least about five yearsafter the initial administration.
 13. The anti-PD-1 antibody for useaccording to any one of claims 1 to 12, wherein the subject has a tumorthat has ≥1% PD-L1 expression.
 14. The anti-PD-1 antibody for useaccording to any one of claims 1 to 13, wherein the subject has a tumorthat has ≥5% PD-L1 expression.
 15. A kit comprising: (a) a dosageranging from about 4 mg to about 500 mg of an antibody or anantigen-binding portion thereof that binds specifically to a ProgrammedDeath-1 (PD-1) receptor and inhibits PD-1 activity (“an anti-PD-1antibody”); (b) a dosage ranging from about 4 mg to about 500 mg of anantibody or an antigen-binding portion thereof that binds specificallyto Cytotoxic T-Lymphocyte Antigen-4 (CTLA-4) and inhibits CTLA-4activity (“a CTLA-4 antibody”); and (c) instructions for using the PD-1antibody and the CTLA-4 antibody according to any one of claims 1 to 14.